Tumor suppression and inflammation: controlling the senescence associated se...adamfreund
This is the powerpoint presentation from a talk I gave at a conference in October, 2009. It will be hard to follow without the spoken part, but it will hopefully give anyone who is interested a brief introduction to my thesis research.
CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes Jaehee Jeong
This document discusses a study that uses CRISPR/Cas9 gene editing on human tripronuclear (3PN) zygotes. 3PN zygotes contain one egg nucleus and two sperm nuclei and are typically discarded. The study aims to edit the HBB gene in the zygotes using CRISPR/Cas9. It finds a 52% cleavage efficiency and some evidence of gene editing, demonstrating the potential of using 3PN zygotes to study CRISPR/Cas9 gene editing in early human development while avoiding ethical issues with normal embryos. However, further improvements are still needed to increase specificity and accuracy before clinical applications.
1. Researchers used CRISPR/Cas9 to efficiently generate biallelic RAG1 knockout in mouse embryonic stem cells. They designed single-guide RNAs targeting RAG1 and transfected stem cells with Cas9, achieving indels in 92% of clones, including 59% with homozygous out-of-frame mutations.
2. The RAG1 knockout stem cell lines maintained pluripotent gene expression and normal morphology. CRISPR/Cas9 allowed faster generation of RAG1 knockout mice than previous methods by creating chimeric embryos.
3. Precisely designed single-guide RNAs and targeting multiple sites simultaneously enhanced CRISPR/Cas9's ability to introduce double-
1. Dr. Madhuri Hegde presented on detecting small intragenic deletions using targeted comparative genomic hybridization (aCGH).
2. She discussed several examples of deletions less than 2.5 kb detected by aCGH in disease genes including PAH, STK11, HPRT1, and EMD.
3. She concluded that aCGH is a valuable tool for detecting small intragenic deletions and providing insights into deletion mechanisms.
This document summarizes molecular pathology of pre-mRNA splicing and discusses how mutations can affect splicing, leading to disease. It describes various methods used to identify pathological splicing mutations experimentally, including RT-PCR to detect mutations. It also discusses how RNA secondary structure and bioinformatics tools can influence splicing and be used to predict structure. Finally, it outlines emerging therapies targeting pre-mRNA splicing, such as antisense oligonucleotides, to treat diseases.
This document discusses genetic polymorphisms and their role in disease pathogenesis and complications. It begins with definitions of health and disease, then discusses various disease causative agents including environmental, genetic, and microbial factors. It describes the organization of the human genome and genes. The document outlines DNA structure and organization, including DNA packaging into chromosomes. It discusses polymorphisms, mutations, and how they can be analyzed using restriction endonucleases and techniques like PCR and gel electrophoresis. Specific examples are provided of how genetic variations can impact disease by changing enzymes, receptors, transport proteins, and other molecular factors.
This document discusses gene editing applications using CRISPR-Cas9, including in gametes and embryos. It provides background on the development of CRISPR-Cas9 as a gene editing tool. Genome editing has been applied to male and female germ cells in animal models and research embryos to correct genetic mutations. However, human embryo genome editing faces limitations such as mosaicism and off-target effects. While genome editing holds promise for treating genetic diseases, more research is needed to improve specificity and fidelity before clinical applications.
Recombinant protein expression and purification Lecturetest
The document discusses recombinant protein expression and engineering. It describes:
1) Cloning or synthesizing the gene of interest, making an expression construct, transfecting cells, purifying the recombinant protein.
2) Factors to consider like the protein's origin (prokaryotic/eukaryotic), required post-translational modifications, and available expression systems.
3) A case study expressing recombinant human alpha-1-acid glycoprotein in E. coli, including vector construction, periplasmic extraction, affinity purification, and yield.
Tumor suppression and inflammation: controlling the senescence associated se...adamfreund
This is the powerpoint presentation from a talk I gave at a conference in October, 2009. It will be hard to follow without the spoken part, but it will hopefully give anyone who is interested a brief introduction to my thesis research.
CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes Jaehee Jeong
This document discusses a study that uses CRISPR/Cas9 gene editing on human tripronuclear (3PN) zygotes. 3PN zygotes contain one egg nucleus and two sperm nuclei and are typically discarded. The study aims to edit the HBB gene in the zygotes using CRISPR/Cas9. It finds a 52% cleavage efficiency and some evidence of gene editing, demonstrating the potential of using 3PN zygotes to study CRISPR/Cas9 gene editing in early human development while avoiding ethical issues with normal embryos. However, further improvements are still needed to increase specificity and accuracy before clinical applications.
1. Researchers used CRISPR/Cas9 to efficiently generate biallelic RAG1 knockout in mouse embryonic stem cells. They designed single-guide RNAs targeting RAG1 and transfected stem cells with Cas9, achieving indels in 92% of clones, including 59% with homozygous out-of-frame mutations.
2. The RAG1 knockout stem cell lines maintained pluripotent gene expression and normal morphology. CRISPR/Cas9 allowed faster generation of RAG1 knockout mice than previous methods by creating chimeric embryos.
3. Precisely designed single-guide RNAs and targeting multiple sites simultaneously enhanced CRISPR/Cas9's ability to introduce double-
1. Dr. Madhuri Hegde presented on detecting small intragenic deletions using targeted comparative genomic hybridization (aCGH).
2. She discussed several examples of deletions less than 2.5 kb detected by aCGH in disease genes including PAH, STK11, HPRT1, and EMD.
3. She concluded that aCGH is a valuable tool for detecting small intragenic deletions and providing insights into deletion mechanisms.
This document summarizes molecular pathology of pre-mRNA splicing and discusses how mutations can affect splicing, leading to disease. It describes various methods used to identify pathological splicing mutations experimentally, including RT-PCR to detect mutations. It also discusses how RNA secondary structure and bioinformatics tools can influence splicing and be used to predict structure. Finally, it outlines emerging therapies targeting pre-mRNA splicing, such as antisense oligonucleotides, to treat diseases.
This document discusses genetic polymorphisms and their role in disease pathogenesis and complications. It begins with definitions of health and disease, then discusses various disease causative agents including environmental, genetic, and microbial factors. It describes the organization of the human genome and genes. The document outlines DNA structure and organization, including DNA packaging into chromosomes. It discusses polymorphisms, mutations, and how they can be analyzed using restriction endonucleases and techniques like PCR and gel electrophoresis. Specific examples are provided of how genetic variations can impact disease by changing enzymes, receptors, transport proteins, and other molecular factors.
This document discusses gene editing applications using CRISPR-Cas9, including in gametes and embryos. It provides background on the development of CRISPR-Cas9 as a gene editing tool. Genome editing has been applied to male and female germ cells in animal models and research embryos to correct genetic mutations. However, human embryo genome editing faces limitations such as mosaicism and off-target effects. While genome editing holds promise for treating genetic diseases, more research is needed to improve specificity and fidelity before clinical applications.
Recombinant protein expression and purification Lecturetest
The document discusses recombinant protein expression and engineering. It describes:
1) Cloning or synthesizing the gene of interest, making an expression construct, transfecting cells, purifying the recombinant protein.
2) Factors to consider like the protein's origin (prokaryotic/eukaryotic), required post-translational modifications, and available expression systems.
3) A case study expressing recombinant human alpha-1-acid glycoprotein in E. coli, including vector construction, periplasmic extraction, affinity purification, and yield.
This document provides information about epigenetic profiling and biomarkers. It discusses a lab for bioinformatics and computational genomics that has over 100 employees working on topics like genome hacking, hardware engineering, and molecular biology. It introduces epigenetics and how changes in gene expression and phenotype can occur without changes to the DNA sequence. Examples are provided of how DNA methylation differs between stem cell types and how reprogramming the methylome can occur. The potential role of epigenetics in cancer is discussed, along with examples of methylation biomarkers like MGMT for glioblastoma. The document describes next generation epigenetic profiling techniques like MBD-Seq and how deep sequencing can study methylation heterogeneity. It outlines efforts to integrate
dkNET Webinar - nPOD Nanotomy: Large-Scale Electron Microscopy Database For H...dkNET
Abstract
Imaging of macromolecules and organelles in the context of cells and tissues is challenging because of the different scales and big data sharing. High resolution imaging of ultrastructure using electron microscopy (EM) typically has a small field of view. Panorama EM views, which we name nanotomy (nano-anatomy), now cross orders of magnitude scales (http://www.nanotomy.org). The open-source sharing allows reuse of data for further analysis, e.g. of structures that were not the focus of the primary study. Nanotomy will likely become the future standard routine EM technique for tissue and cell imaging. In this talk I will highlight the technique and the recent database of nanotomy of human pancreas tissue obtained from the Network for Pancreatic Organ donors with Diabetes.
Research: cellbiology.nl
lEM-dbase: nanotomy.org
UMIC core: umic.info
Presenter: Ben N. G. Giepmans, PhD, Associate Professor, Biomedical Sciences of Cells & Systems, UMC Groningen, The Netherlands
Upcoming webinars schedule: https://dknet.org/about/webinar
CRISPR-Cas9 is a genome editing technique that allows DNA to be precisely cut and modified. It involves using the Cas9 enzyme, guided by RNA, to cut DNA at a specific target location. The cell's DNA repair machinery can then introduce changes by adding, removing, or replacing DNA segments. CRISPR-Cas9 was adapted from a natural defense system in bacteria against viruses. It holds promise for treating genetic diseases but also raises ethical concerns about editing human embryos or germline cells.
This document provides an overview of CRISPR/Cas9 genome editing. It discusses the history and limitations of prior genome engineering techniques like recombinant DNA and zinc finger nucleases. It then explains how CRISPR/Cas9 works as a RNA-guided DNA endonuclease and how this allows it to efficiently and specifically edit genomes. The document outlines several applications of CRISPR/Cas9 like generating knockout animals and cell lines. It also notes some concerns about using the technique for human genome editing.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying the genomes of organisms ranging from E. coli to humans. In this presentation, we discuss various methods for generating the crRNA and tracrRNA components that are required for guiding the Cas9 endonuclease to genomic targets. You will also learn how to optimize a new 2-part CRISPR RNA system from IDT that offers multiple benefits over other technologies.
Western blotting is a technique used to detect specific proteins in a sample containing a mixture of proteins. It involves separating proteins by electrophoresis, transferring them to a membrane, and using antibodies to identify a specific protein target through immunodetection. Key steps include extracting proteins, separating by SDS-PAGE gel electrophoresis, transferring to a membrane, blocking the membrane, incubating with primary and secondary antibodies, and detecting the target protein band using an enzyme substrate reaction. It is a widely used analytical technique in cell and molecular biology research.
CRISPR/Cas9 gene editing is based on a microbial restriction system, that has been harnessed for genome targeting using only a short sequence of RNA as a guide.
The beauty of the system is that unlike protein binding based technologies such as Zinc Fingers and TALENs which require complex protein engineering, the design rules are very simple, and it is this fact that is allowing CRISPR to take genome engineering from a relatively niche persuit to the mainstream scientific community.
The principle of the system is that a short guide RNA, homologous to the target site recruits a nuclease – Cas9
This then cuts the dsDNA, triggering repair by either the low fidelity NHEJ pathway, or by HDR in the presence of an exogenous donor sequence.
High Efficiencies for both knockouts and knock-ins have been reported and whilst there are understandable concerns about specificity, new methodologies to address these are now being developed
The system itself is comprised of three key components
the Cas9 protein, which cuts/cleaves the DNA and
Two RNAs - a crispr RNA contains the sequence homologous to the target site and a trans-activating crisprRNA (or TracrRNA) which recruits the nuclease/crispr complex
For genome editing, the crisperRNA and TraceRNA are generally now constructed together into a single guideRNA or sgRNA
Genome editing is elicited through hybridization of the sgRNA with its matching genomic sequence, and the recruitment of the Cas9, which cleaves at the target site.
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...AvactaLifeSciences
Avacta Life Sciences Exhibits Affimers at Global Protein Engineering Summit
Avacta Life Sciences exhibited recently at the Global Protein Engineering Summit ("PEGS") where it presented its Affimer technology.
You can read more about Affimer technology here http://www.avactalifesciences.com
PEGS is considered to be the essential protein engineering meeting where commercial and academic progress in protein engineering is showcased and this year it attracted over 1800 delegates from across the globe to Boston. Avacta Life Sciences presented its Affimer technology for the first time at a PEGS meeting with technical exhibits and a presentation by the CSO, Paul Ko Ferrigno, entitled "Biological Recognition: Beyond the Antibody."*
The exhibition booth was busy with over 80 delegates talking to the Avacta Life Science management team over the four days of the summit. The feedback on the Affimer technology was very positive, in particular, the short development times and excellent stability were highlighted by delegates as key advantages of Affimers over antibodies. There was also a strong interest in Affimers from the management of companies developing biological therapeutics who were keen to learn more about the potential of Affimers as novel therapeutics.
In addition, several companies were interested in the use of Affimers as an alternative to antibodies in diagnostic devices, mainly because they could generate binders against new biomarkers much more quickly and evaluate them in higher numbers.
The benefits of Affimer microarrays for biomarker discovery also resonated with diagnostic developers who appreciated the advantage of being able to evaluate significantly larger numbers of potential biomarkers more cost and time effectively than by mass spectrometry. The potential of the arrays for multiplexed solutions for clinical diagnosis and monitoring during drug trials was also something that generated interest amongst those delegates.
Matt Johnson, Chief Technical Officer of Avacta Life Sciences commented: "It was great to experience face to face the level of interest in Affimers. The majority of people I spoke to were either having problems raising antibodies to their target of interest or just couldn't use antibodies because of the type of assays they wanted to perform. Many of the presentations focused around the use of antibody fragments for intra-cellular studies which is a rapidly growing area that holds great interest for drug and diagnostics developers. It is an area where there are clear advantages for Affimers over antibody fragments which don't behave well in the cytoplasm.
"The general enthusiasm around Affimers was very encouraging and the amount of interest generated by the potential of Affimers as therapeutics and by the Affimer arrays for biomarker discovery only reinforces my excitement around this new technology."
CRISPR system is very simple, consisted of a Cas9 protein and a single guided RNA. With the guidance of sgRNA, Cas9 could cause a double stranded breaks in the target site.
CRISPR/Cas9 system consists of a “guide” RNA (gRNA) and a bacterial CRISPR-associated endonuclease (Cas9). The gRNA is a short synthetic RNA composed of a Cas9-binding “scaffold” sequence and ∼20 nucleotide “targeting” sequence that defines the target genomic site to be modified.
https://www.creative-biogene.com/Services/Stable-cell-line-generation/Custom-Genome-Editing-Cell-Lines.html
The document discusses the CRISPR/Cas9 system. It describes how CRISPR/Cas9 uses a Cas9 protein guided by a single guide RNA to recognize and cut target DNA. The system has three stages: adaptation, expression and processing of CRISPR RNA, and interference where the Cas9 protein complex cuts the target DNA. CRISPR/Cas9 can be engineered to act as a nuclease, nickase, or inactive dead Cas9 for gene regulation applications like activation or repression. It provides a gift from nature for precise genome editing and regulation.
Recent breakthroughs in genome editing technology have led to a rapid adoption that parallels that seen with RNAi. And like RNAi, these methods are taking the scientific world by storm, with high profile publications in fields as diverse as HIV treatment, stem cell therapy, food crop modification and drug development to name but a few.
Critically, the endogenous modification of genes enables the study of their function in a physiological context. It also overcomes some of the artefacts that can result from established techniques such as transgenesis and RNAi, which have mislead researchers with false positives or negatives. Until recently however genome editing required considerable technical expertise, and consequently was a relatively niche pursuit.
In this talk we will look at how the latest developments in genome editing tools have changed this, with improvements in both ease-of-use and targeting efficiency, as well as a concomitant reduction in costs opening up these approaches to the wider scientific community.
Rapid adoption of the CRISPR/Cas9 system has for example led to a long list of organisms and tissues in which genetic changes have been made with high efficiency. Other technologies such as recombinant adeno-associated virus (rAAV) offer further precision, stimulating the cell’s high-fidelity DNA repair pathways to insert exogenous sequence with unrivalled specificity. Targeting efficiency can be improved still further by using the technologies in combination – genome cutting induced by CRISPR can significantly enhance homologous recombination mediated by rAAV.
Despite these rapid advances, some pitfalls remain, and so we’ll discuss some of the key considerations for avoiding these, ranging from simply picking the right tool for the job to designing an experiment that maximises chances of success.
Finally we’ll look at how genome editing is being applied to both basic and translational research, and in both a gene-specific and genome wide manner. For the study of disease associated genes and mutations scientists can now complement wide panels of tumour cells with genetically defined isogenic cell pairs identical in all but precise modifications in their gene of interest. The ease-of-design and efficiency of the CRISPR system is also being exploited for genome wide synthetic lethality screens, facilitating rapid drug target identification with significantly reduced risk of false negatives and off-target false positives. And again, further synergies are achieved when these approaches are combined to look for potential synthetic lethal targets in specific genomic contexts.
This document reviews the use of platelet-rich plasma (PRP) as an enhancement factor for guided bone regeneration and dental implants. It summarizes that PRP delivers concentrated growth factors to sites requiring bone grafting in order to increase bone formation. However, few human studies have been conducted to scientifically validate the efficacy of PRP for bone regeneration. Well-designed controlled studies are still needed to provide evidence that PRP is effective at improving bone regeneration outcomes.
The document discusses several studies related to atherosclerosis and cardiovascular disease:
1) A study finds that a polymorphism in the Fas gene promoter region is a genetic risk factor for myocardial infarction by modulating Fas expression.
2) Immunoglobulin treatment suppresses atherosclerosis in mice via its Fc portion by reducing macrophage accumulation in lesions.
3) Inhibition of NF-kB reduces inflammatory molecule expression and attenuates atherosclerosis in mice.
4) MMP-8 may represent a new collagenolytic pathway in acute plaque disruption based on its levels in carotid plaques from patients.
This document discusses genome editing using the CRISPR-Cas9 system. It begins by introducing three main genome editing technologies - zinc-finger nucleases, TALENs, and the CRISPR-Cas9 system. It then describes the key events in the discovery of CRISPR-Cas9, including its origins as a bacterial defense system. The document outlines the main components of the CRISPR-Cas9 system, including crRNA, tracrRNA, sgRNA, and Cas9. It also summarizes the two main steps in genome editing using CRISPR-Cas9 - knocking out genes and DNA repair. The document concludes by discussing opportunities for applying CRISPR-Cas9 technology across various
Have you considered that protein over-expression or inefficient mRNA knockdown may be masking physiological effects in your assays? Increasingly scientists are moving to endogenous gene-editing to characterise the function of their genes of interest.
Dr Chris Thorne from Cambridge Biotech Horizon Discovery discusses the ground breaking gene-editing technology CRISPR. The simplicity of experimental design has led to rapid adoption of the technology across the scientific community. However, challenges remain.
This Slidedeck focuses specifically on implementing CRISPR experiments, and explore a number of key considerations crucial to maximising chances of targeting success, whether your goal is to generate a knock-out or a knock-in. Chris also takes a look at some of the alternative uses of CRISPR, including sgRNA genome wide synthetic lethality screens.
The slides aim to support those researchers either planning to or already using CRISPR gene-editing in their lab. Horizon Discovery have also recently launched a program aimed specifically at academic cell biologists to promote the adoption of CRISPR by offering FREE CRISPR Reagents for knock-out cell line generation - more information available here. http://www.horizondiscovery.com/what-we-do/discovery-toolbox/genassist-crispr--raav-genome-editing-tools
The Efficiency and Ethics of the CRISPR System in Human EmbryosStephen Cranwell
This document summarizes research on the CRISPR/Cas9 system for genome editing in human embryos. It discusses efforts to understand DNA repair mechanisms after inducing double-strand breaks, reduce off-target mutations, and improve the specificity and efficiency of editing. While the technology shows promise, significant issues around off-target effects, mosaicism, and ethical concerns must still be addressed before any clinical applications. The document concludes that further basic research is needed to advance the field while also having open discussions on societal implications.
1) Wnt/beta-catenin signaling plays a role in skin development and hair follicle cycling. Expression of a truncated beta-catenin in mouse skin leads to de novo hair follicle formation.
2) Mutations in beta-catenin that stabilize the protein are found in 75% of human pilomatricoma tumors, a skin tumor of hair matrix cell origin. This suggests beta-catenin stabilization initiates these tumors.
3) Forced expression of beta-catenin in chick skin can initiate feather bud development and ectopic feather formation by upregulating genes like BMP-2. This supports a role for beta-catenin in initiating skin appendage morphogenesis.
La SCID es una enfermedad genética rara caracterizada por la ausencia de linfocitos T, B y NK debido a un bloqueo en su desarrollo. Puede ser causada por deficiencias en ADA, cadenas gamma, receptores de IL-7 alfa, recombinación de genes o Janus cinasa 3. Los síntomas incluyen infecciones graves y recurrentes desde los primeros meses de vida. El diagnóstico se basa en manifestaciones clínicas e inmunológicas. Los tratamientos son trasplante de médula ósea, ter
This document provides information about epigenetic profiling and biomarkers. It discusses a lab for bioinformatics and computational genomics that has over 100 employees working on topics like genome hacking, hardware engineering, and molecular biology. It introduces epigenetics and how changes in gene expression and phenotype can occur without changes to the DNA sequence. Examples are provided of how DNA methylation differs between stem cell types and how reprogramming the methylome can occur. The potential role of epigenetics in cancer is discussed, along with examples of methylation biomarkers like MGMT for glioblastoma. The document describes next generation epigenetic profiling techniques like MBD-Seq and how deep sequencing can study methylation heterogeneity. It outlines efforts to integrate
dkNET Webinar - nPOD Nanotomy: Large-Scale Electron Microscopy Database For H...dkNET
Abstract
Imaging of macromolecules and organelles in the context of cells and tissues is challenging because of the different scales and big data sharing. High resolution imaging of ultrastructure using electron microscopy (EM) typically has a small field of view. Panorama EM views, which we name nanotomy (nano-anatomy), now cross orders of magnitude scales (http://www.nanotomy.org). The open-source sharing allows reuse of data for further analysis, e.g. of structures that were not the focus of the primary study. Nanotomy will likely become the future standard routine EM technique for tissue and cell imaging. In this talk I will highlight the technique and the recent database of nanotomy of human pancreas tissue obtained from the Network for Pancreatic Organ donors with Diabetes.
Research: cellbiology.nl
lEM-dbase: nanotomy.org
UMIC core: umic.info
Presenter: Ben N. G. Giepmans, PhD, Associate Professor, Biomedical Sciences of Cells & Systems, UMC Groningen, The Netherlands
Upcoming webinars schedule: https://dknet.org/about/webinar
CRISPR-Cas9 is a genome editing technique that allows DNA to be precisely cut and modified. It involves using the Cas9 enzyme, guided by RNA, to cut DNA at a specific target location. The cell's DNA repair machinery can then introduce changes by adding, removing, or replacing DNA segments. CRISPR-Cas9 was adapted from a natural defense system in bacteria against viruses. It holds promise for treating genetic diseases but also raises ethical concerns about editing human embryos or germline cells.
This document provides an overview of CRISPR/Cas9 genome editing. It discusses the history and limitations of prior genome engineering techniques like recombinant DNA and zinc finger nucleases. It then explains how CRISPR/Cas9 works as a RNA-guided DNA endonuclease and how this allows it to efficiently and specifically edit genomes. The document outlines several applications of CRISPR/Cas9 like generating knockout animals and cell lines. It also notes some concerns about using the technique for human genome editing.
The CRISPR/Cas9 system has emerged as one of the leading tools for modifying the genomes of organisms ranging from E. coli to humans. In this presentation, we discuss various methods for generating the crRNA and tracrRNA components that are required for guiding the Cas9 endonuclease to genomic targets. You will also learn how to optimize a new 2-part CRISPR RNA system from IDT that offers multiple benefits over other technologies.
Western blotting is a technique used to detect specific proteins in a sample containing a mixture of proteins. It involves separating proteins by electrophoresis, transferring them to a membrane, and using antibodies to identify a specific protein target through immunodetection. Key steps include extracting proteins, separating by SDS-PAGE gel electrophoresis, transferring to a membrane, blocking the membrane, incubating with primary and secondary antibodies, and detecting the target protein band using an enzyme substrate reaction. It is a widely used analytical technique in cell and molecular biology research.
CRISPR/Cas9 gene editing is based on a microbial restriction system, that has been harnessed for genome targeting using only a short sequence of RNA as a guide.
The beauty of the system is that unlike protein binding based technologies such as Zinc Fingers and TALENs which require complex protein engineering, the design rules are very simple, and it is this fact that is allowing CRISPR to take genome engineering from a relatively niche persuit to the mainstream scientific community.
The principle of the system is that a short guide RNA, homologous to the target site recruits a nuclease – Cas9
This then cuts the dsDNA, triggering repair by either the low fidelity NHEJ pathway, or by HDR in the presence of an exogenous donor sequence.
High Efficiencies for both knockouts and knock-ins have been reported and whilst there are understandable concerns about specificity, new methodologies to address these are now being developed
The system itself is comprised of three key components
the Cas9 protein, which cuts/cleaves the DNA and
Two RNAs - a crispr RNA contains the sequence homologous to the target site and a trans-activating crisprRNA (or TracrRNA) which recruits the nuclease/crispr complex
For genome editing, the crisperRNA and TraceRNA are generally now constructed together into a single guideRNA or sgRNA
Genome editing is elicited through hybridization of the sgRNA with its matching genomic sequence, and the recruitment of the Cas9, which cleaves at the target site.
Avacta Life Sciences Affimers Presentation Global Protein Engineering Summit ...AvactaLifeSciences
Avacta Life Sciences Exhibits Affimers at Global Protein Engineering Summit
Avacta Life Sciences exhibited recently at the Global Protein Engineering Summit ("PEGS") where it presented its Affimer technology.
You can read more about Affimer technology here http://www.avactalifesciences.com
PEGS is considered to be the essential protein engineering meeting where commercial and academic progress in protein engineering is showcased and this year it attracted over 1800 delegates from across the globe to Boston. Avacta Life Sciences presented its Affimer technology for the first time at a PEGS meeting with technical exhibits and a presentation by the CSO, Paul Ko Ferrigno, entitled "Biological Recognition: Beyond the Antibody."*
The exhibition booth was busy with over 80 delegates talking to the Avacta Life Science management team over the four days of the summit. The feedback on the Affimer technology was very positive, in particular, the short development times and excellent stability were highlighted by delegates as key advantages of Affimers over antibodies. There was also a strong interest in Affimers from the management of companies developing biological therapeutics who were keen to learn more about the potential of Affimers as novel therapeutics.
In addition, several companies were interested in the use of Affimers as an alternative to antibodies in diagnostic devices, mainly because they could generate binders against new biomarkers much more quickly and evaluate them in higher numbers.
The benefits of Affimer microarrays for biomarker discovery also resonated with diagnostic developers who appreciated the advantage of being able to evaluate significantly larger numbers of potential biomarkers more cost and time effectively than by mass spectrometry. The potential of the arrays for multiplexed solutions for clinical diagnosis and monitoring during drug trials was also something that generated interest amongst those delegates.
Matt Johnson, Chief Technical Officer of Avacta Life Sciences commented: "It was great to experience face to face the level of interest in Affimers. The majority of people I spoke to were either having problems raising antibodies to their target of interest or just couldn't use antibodies because of the type of assays they wanted to perform. Many of the presentations focused around the use of antibody fragments for intra-cellular studies which is a rapidly growing area that holds great interest for drug and diagnostics developers. It is an area where there are clear advantages for Affimers over antibody fragments which don't behave well in the cytoplasm.
"The general enthusiasm around Affimers was very encouraging and the amount of interest generated by the potential of Affimers as therapeutics and by the Affimer arrays for biomarker discovery only reinforces my excitement around this new technology."
CRISPR system is very simple, consisted of a Cas9 protein and a single guided RNA. With the guidance of sgRNA, Cas9 could cause a double stranded breaks in the target site.
CRISPR/Cas9 system consists of a “guide” RNA (gRNA) and a bacterial CRISPR-associated endonuclease (Cas9). The gRNA is a short synthetic RNA composed of a Cas9-binding “scaffold” sequence and ∼20 nucleotide “targeting” sequence that defines the target genomic site to be modified.
https://www.creative-biogene.com/Services/Stable-cell-line-generation/Custom-Genome-Editing-Cell-Lines.html
The document discusses the CRISPR/Cas9 system. It describes how CRISPR/Cas9 uses a Cas9 protein guided by a single guide RNA to recognize and cut target DNA. The system has three stages: adaptation, expression and processing of CRISPR RNA, and interference where the Cas9 protein complex cuts the target DNA. CRISPR/Cas9 can be engineered to act as a nuclease, nickase, or inactive dead Cas9 for gene regulation applications like activation or repression. It provides a gift from nature for precise genome editing and regulation.
Recent breakthroughs in genome editing technology have led to a rapid adoption that parallels that seen with RNAi. And like RNAi, these methods are taking the scientific world by storm, with high profile publications in fields as diverse as HIV treatment, stem cell therapy, food crop modification and drug development to name but a few.
Critically, the endogenous modification of genes enables the study of their function in a physiological context. It also overcomes some of the artefacts that can result from established techniques such as transgenesis and RNAi, which have mislead researchers with false positives or negatives. Until recently however genome editing required considerable technical expertise, and consequently was a relatively niche pursuit.
In this talk we will look at how the latest developments in genome editing tools have changed this, with improvements in both ease-of-use and targeting efficiency, as well as a concomitant reduction in costs opening up these approaches to the wider scientific community.
Rapid adoption of the CRISPR/Cas9 system has for example led to a long list of organisms and tissues in which genetic changes have been made with high efficiency. Other technologies such as recombinant adeno-associated virus (rAAV) offer further precision, stimulating the cell’s high-fidelity DNA repair pathways to insert exogenous sequence with unrivalled specificity. Targeting efficiency can be improved still further by using the technologies in combination – genome cutting induced by CRISPR can significantly enhance homologous recombination mediated by rAAV.
Despite these rapid advances, some pitfalls remain, and so we’ll discuss some of the key considerations for avoiding these, ranging from simply picking the right tool for the job to designing an experiment that maximises chances of success.
Finally we’ll look at how genome editing is being applied to both basic and translational research, and in both a gene-specific and genome wide manner. For the study of disease associated genes and mutations scientists can now complement wide panels of tumour cells with genetically defined isogenic cell pairs identical in all but precise modifications in their gene of interest. The ease-of-design and efficiency of the CRISPR system is also being exploited for genome wide synthetic lethality screens, facilitating rapid drug target identification with significantly reduced risk of false negatives and off-target false positives. And again, further synergies are achieved when these approaches are combined to look for potential synthetic lethal targets in specific genomic contexts.
This document reviews the use of platelet-rich plasma (PRP) as an enhancement factor for guided bone regeneration and dental implants. It summarizes that PRP delivers concentrated growth factors to sites requiring bone grafting in order to increase bone formation. However, few human studies have been conducted to scientifically validate the efficacy of PRP for bone regeneration. Well-designed controlled studies are still needed to provide evidence that PRP is effective at improving bone regeneration outcomes.
The document discusses several studies related to atherosclerosis and cardiovascular disease:
1) A study finds that a polymorphism in the Fas gene promoter region is a genetic risk factor for myocardial infarction by modulating Fas expression.
2) Immunoglobulin treatment suppresses atherosclerosis in mice via its Fc portion by reducing macrophage accumulation in lesions.
3) Inhibition of NF-kB reduces inflammatory molecule expression and attenuates atherosclerosis in mice.
4) MMP-8 may represent a new collagenolytic pathway in acute plaque disruption based on its levels in carotid plaques from patients.
This document discusses genome editing using the CRISPR-Cas9 system. It begins by introducing three main genome editing technologies - zinc-finger nucleases, TALENs, and the CRISPR-Cas9 system. It then describes the key events in the discovery of CRISPR-Cas9, including its origins as a bacterial defense system. The document outlines the main components of the CRISPR-Cas9 system, including crRNA, tracrRNA, sgRNA, and Cas9. It also summarizes the two main steps in genome editing using CRISPR-Cas9 - knocking out genes and DNA repair. The document concludes by discussing opportunities for applying CRISPR-Cas9 technology across various
Have you considered that protein over-expression or inefficient mRNA knockdown may be masking physiological effects in your assays? Increasingly scientists are moving to endogenous gene-editing to characterise the function of their genes of interest.
Dr Chris Thorne from Cambridge Biotech Horizon Discovery discusses the ground breaking gene-editing technology CRISPR. The simplicity of experimental design has led to rapid adoption of the technology across the scientific community. However, challenges remain.
This Slidedeck focuses specifically on implementing CRISPR experiments, and explore a number of key considerations crucial to maximising chances of targeting success, whether your goal is to generate a knock-out or a knock-in. Chris also takes a look at some of the alternative uses of CRISPR, including sgRNA genome wide synthetic lethality screens.
The slides aim to support those researchers either planning to or already using CRISPR gene-editing in their lab. Horizon Discovery have also recently launched a program aimed specifically at academic cell biologists to promote the adoption of CRISPR by offering FREE CRISPR Reagents for knock-out cell line generation - more information available here. http://www.horizondiscovery.com/what-we-do/discovery-toolbox/genassist-crispr--raav-genome-editing-tools
The Efficiency and Ethics of the CRISPR System in Human EmbryosStephen Cranwell
This document summarizes research on the CRISPR/Cas9 system for genome editing in human embryos. It discusses efforts to understand DNA repair mechanisms after inducing double-strand breaks, reduce off-target mutations, and improve the specificity and efficiency of editing. While the technology shows promise, significant issues around off-target effects, mosaicism, and ethical concerns must still be addressed before any clinical applications. The document concludes that further basic research is needed to advance the field while also having open discussions on societal implications.
1) Wnt/beta-catenin signaling plays a role in skin development and hair follicle cycling. Expression of a truncated beta-catenin in mouse skin leads to de novo hair follicle formation.
2) Mutations in beta-catenin that stabilize the protein are found in 75% of human pilomatricoma tumors, a skin tumor of hair matrix cell origin. This suggests beta-catenin stabilization initiates these tumors.
3) Forced expression of beta-catenin in chick skin can initiate feather bud development and ectopic feather formation by upregulating genes like BMP-2. This supports a role for beta-catenin in initiating skin appendage morphogenesis.
La SCID es una enfermedad genética rara caracterizada por la ausencia de linfocitos T, B y NK debido a un bloqueo en su desarrollo. Puede ser causada por deficiencias en ADA, cadenas gamma, receptores de IL-7 alfa, recombinación de genes o Janus cinasa 3. Los síntomas incluyen infecciones graves y recurrentes desde los primeros meses de vida. El diagnóstico se basa en manifestaciones clínicas e inmunológicas. Los tratamientos son trasplante de médula ósea, ter
Severe combined immunodeficiency (SCID) is a genetic disorder characterized by the absence of functional T lymphocytes and B lymphocytes, resulting in impaired adaptive immune system. There are several known types of SCID caused by mutations in different genes. The most common treatment is bone marrow transplantation, with success seen in transplants done in early infancy. Gene therapy is also being explored as a potential treatment through inserting missing genes into hematopoietic stem cells, though past trials increased leukemia risk and more research is still needed.
Severe combined immunodeficiency (SCID), also known as "Bubble Boy Disease", is a primary immunodeficiency caused by mutations in several genes resulting in the lack of T-cells and sometimes B-cells and NK cells as well. It was widely known due to David Vetter who lived in a germ-free plastic bubble for 12 years. SCID is diagnosed through newborn screening, blood tests, and showing a lack of white blood cells in newborns. Treatments include supportive therapies like antibiotics and immunoglobulin injections as well as curative therapies such as stem cell transplant or gene therapy.
This document discusses severe combined immunodeficiency (SCID), a genetic disorder characterized by defective development of functional T and B cells. The document covers the causes of SCID including mutations that affect cytokine receptors, enzymes, and genes involved in lymphocyte development. Signs and symptoms of SCID are described as well as diagnostic tests including low T cell counts and lack of response to mitogens. Treatment options for SCID such as hematopoietic stem cell transplantation and gene therapy are also mentioned.
Introduction, causes and symptoms, Mechanism and treatment are been explained about this deadly disease SCID where production of T and B cells is affected.
Severe combined immunodeficiency (SCID), also known as bubble boy disease, is a genetic disorder where both the B cell and T cell arms of the immune system are impaired. There are several types of SCID resulting from defects in different genes. Symptoms include life-threatening infections from an early age. Treatment involves preventing infections, enzyme therapy, gene therapy, or bone marrow transplant from a matched or half-matched donor to rebuild the immune system. Transplants have been successful but carry risks, as seen in the original "bubble boy" who died from a virus in his transplanted marrow.
2013 05 BEA - ’Mobile is eating the World’Benedict Evans
Mobile devices such as smartphones and tablets are dramatically increasing in sales and dominating the consumer technology market. Global PC sales peaked in 2011 and have declined since, being overtaken by surging tablet sales. By 2017 there will be over 7 billion mobile subscribers, compared to just over 1 billion PC users. This fundamental shift is changing industries like media, advertising, and retail as mobile internet usage grows and mobile platforms like Apple's iOS and Google's Android come to dominate the market.
This document discusses cisgenesis and intragenesis, which involve genetically modifying a crop plant using genes isolated from a crossable donor plant or from the same plant species, respectively. It defines cisgenic and intragenic plants and outlines their similarities and differences. It describes the prerequisites and various methods for constructing intragenic vectors and producing marker-free cisgenic/intragenic plants. The document presents several case studies demonstrating the development and evaluation of cisgenic plants with improved disease resistance. It discusses regulations around cisgenic/intragenic crops in different countries and potential benefits compared to transgenic and conventional breeding approaches.
This document discusses the CRISPR-Cas9 genome editing technique. It begins with an overview of genome editing and provides a brief history. It then focuses on explaining CRISPR-Cas9, including its key components, how it was discovered as a natural bacterial immune system, and how it functions as a genomic tool. The document outlines the general CRISPR-Cas9 protocol and recent advances in the technique. It discusses applications in agriculture and for diseases. It also touches on advantages and limitations, as well as ethical issues. Two case studies are provided that demonstrate using CRISPR-Cas9 to modify genes in rice plants.
1. The ChampionChIP system allows for epigenetic analysis of histone modifications and transcription factor binding in a single day. It uses validated antibodies, primers, and qPCR arrays to analyze multiple genomic regions simultaneously.
2. The system was used to analyze dynamic bivalent histone modification patterns of pluripotency genes during mouse embryonic carcinoma cell differentiation induced by retinoic acid. Distinct patterns were observed for genes and heterochromatic regions.
3. The system correctly identified histone modification distribution and was used to map modifications around the CDKN1A gene. It also analyzed p53 binding and correlated gene expression changes in response to drug treatment in cancer cell lines.
Los días 20 y 21 de octubre de 2016, la Fundacion Ramón Areces organizó un simposio internacional para analizar las 'Enfermedades raras de la piel: de la clínica al gen y viceversa'. El doctor Fernando Larcher Laguzzi, del CIEMAT-Universidad Carlos III de Madrid-IIS Fundación Jiménez Díaz, ejerció de coordinador.
CRISPR- Trap: a clean approach for the generation of gene knockouts and gene replacements in human cells.- a paper is taken for lab presentation. A very good technique having advantages over conventional KO approaches and allow for the generation of clean CRISPR/ Cas9- based KOs.
Ø Researchers used CRISPR/Cas9 nuclease and nickase to target and repair the c.456+4A>T mutation in the FANCC gene, which causes Fanconi anemia (FA), in human fibroblasts derived from an FA patient.
Ø Both nuclease and nickase mediated repair of the mutation, restoring normal FANCC gene function, though nickase was more efficient due to preferentially inducing the homology-directed repair pathway.
Ø Genome-wide analyses found no off-target effects, confirming the CRISPR reagents specifically targeted the intended FANCC locus. This provides proof-of-principle that CR
CRISPR/Cas9 is a powerful genome editing tool that allows genetic material to be added, altered or removed at specific locations in the genome. It involves a bacterial adaptive immune system where CRISPR sequences and Cas genes work together. The Cas9 protein uses a guide RNA to introduce double stranded breaks at targeted DNA sequences. This enables precise genome editing through non-homologous end joining or homology directed repair. CRISPR/Cas9 provides a simple and accurate way to modify genes for applications in research, medicine, agriculture and more. While it holds great promise, there are also limitations and concerns regarding off-target effects that researchers continue working to address.
A new system called the "admid system" has been developed for efficiently generating recombinant adenoviruses in E. coli. The system uses Tn7-mediated transposition to insert expression cassettes into an adenoviral genome plasmid (admid) maintained in E. coli. Transfer vectors containing the expression cassette flanked by Tn7 elements transpose the cassette into the E1 region of the admid. Recombinant admids produce infectious adenovirus after transfection into producer cells. The system generates pure, clonal adenovirus stocks without multiple rounds of purification. It allows rapid, high-throughput production of recombinant adenoviruses for gene therapy and other applications.
Speaker: Benedict C. S. Cross, PhD, Team leader (Discovery Screening), Horizon Discovery
CRISPR–Cas9 mediated genome editing provides a highly efficient way to probe gene function. Using this technology, thousands of genes can be knocked out and their function assessed in a single experiment. We have conducted over 150 of these complex and powerful screens and will use our experience to guide you through the process of screen design, performance and analysis.
We'll be discussing:
• How to use CRISPR screening for target ID and validation, understanding drug MOA and patient stratification
• The screen design, quality control and how to evaluate success of your screening program
• Horizon’s latest developments to the platform
• Horizon’s novel approaches to target validation screening
This document summarizes a presentation on using CRISPR-Cas9 for crop improvement. It begins with an introduction to CRISPR-Cas9 and how it is used to edit genomes by removing, adding, or altering DNA sequences. It then discusses the mechanism of the CRISPR-Cas9 complex and how it creates breaks in DNA that are repaired. The document reviews several case studies where CRISPR was used to modify crops, including creating low-gluten wheat and improving rice. It finds that CRISPR can efficiently edit multiple genes simultaneously with few off-target effects. The conclusion states that CRISPR is revolutionizing agriculture by enabling the creation of higher yielding, more resistant crop varieties without transgenes.
This document summarizes information about the CRISPR Cas9 genome editing tool. It discusses how CRISPR Cas9 uses guide RNA and the Cas9 enzyme to create targeted double-strand breaks in DNA, allowing genes to be knocked out or altered. The document outlines the history and mechanism of CRISPR Cas9, compares it to other genome editing tools, discusses its applications in plant breeding including reducing off-target effects, and provides an example of using it to create parthenocarpic tomato plants.
The document describes a new reporter system for evaluating the specificity and efficacy of CRISPR/Cas9 and guide RNAs (gRNAs). The system uses an EGFP reporter plasmid containing the target sequence of interest. When co-transfected with Cas9 and the corresponding gRNA, double strand breaks at the target site disrupt EGFP expression. This allows unbiased measurement of Cas9 and gRNA activity levels. The document demonstrates that mismatches between gRNAs and target sequences can occur anywhere in the gRNA and depend on both the gRNA sequence and Cas9 construct used. This reporter system promises to improve genomic engineering success rates by facilitating selection of optimal gRNA and Cas9 combinations.
CRISPR theory mechanism and applications || كرسبر النظريه وطريقه العمل والتطب...Mohemmad Osama
CRISPR-Cas9 is a gene editing technique that allows DNA to be added, removed, or altered at specific locations in the genome. It involves using the Cas9 protein to cut DNA at a targeted site, which can be programmed by changing a short RNA sequence. CRISPR is simpler and easier than previous gene editing methods. It has enabled unprecedented efficiency and ease of use for gene therapy applications in correcting genetic defects and treating diseases. However, its rapid development has also led to an intensifying patent war and debate over its ethical use.
This study aimed to optimize the CRISPR/Cas9 genome editing protocol for efficient homozygous gene knock-in in human induced pluripotent stem cells (iPSCs). The researchers targeted the CD90 locus for replacement with the mouse ortholog Cd90 and tested various experimental conditions. After optimization, CRISPR efficiency increased from 0.28% to 11.8% homozygous knock-in as determined by flow cytometry. Key conditions implicated in higher efficiency included plasmid concentrations and quality, Cas9 delivery method, nucleofection device, recovery conditions, and cell concentration during nucleofection.
This document discusses next generation sequencing (NGS) and its applications in preimplantation genetic diagnosis (PGD). It describes how NGS can simultaneously detect chromosome abnormalities and gene defects in single cells. Studies show NGS has the same accuracy as array comparative genomic hybridization for detecting aneuploidy, and can also detect mutations. NGS has been successfully used to analyze blastocysts and single cells, identifying euploid and aneuploid embryos as well as specific gene mutations. This makes NGS useful for PGD to evaluate chromosome number and identify genetic disorders in embryos prior to implantation.
This document discusses next generation sequencing (NGS) and its applications in preimplantation genetic diagnosis (PGD). It describes how NGS can simultaneously detect chromosome abnormalities and gene defects in single cells. Studies show NGS has the same accuracy as array comparative genomic hybridization for detecting aneuploidy, and can also detect mutations. NGS has been successfully used to analyze blastocysts and single cells to identify euploid and aneuploid embryos as well as specific gene mutations. This makes NGS useful for PGD to select embryos without chromosome issues or gene defects.
In Situ Polymerase Chain Reaction (In situ PCR) is a powerful method that detects minute quantities of rare or single-copy number nucleic acid sequences in frozen or paraffin-embedded cells or tissue sections for the localization of those sequences within the cells. The principle of this method involves tissue fixing (to preserve the cell morphology) and subsequent treatment with proteolytic digestion (to provide access for the PCR reagents to the target DNA). The target sequences are amplified by those reagents and then detected by standard immunocytochemical protocols. In situ PCR combines the sensitivity of PCR or RT-PCR amplification along with the ability to perform morphological analysis on the same sample, and thus it is an attractive tool in diagnostic applications. One of the most prominent applications is the detection of infectious disease agents including HIV-1, HBV, HPV, HHV-6, CMV, and EBV.
This document discusses various approaches for therapy of genetic diseases, including conventional and gene therapy approaches. Conventional approaches include dietary therapy, protein/enzyme replacement, pharmacal therapy, and surgery. Gene therapy involves the deliberate introduction of genetic material into human cells for therapeutic purposes, and can be done through somatic or germline cell gene therapy, as well as ex vivo or in vivo approaches. Viral vectors are commonly used to deliver the therapeutic gene to target cells.
Applied StemCell Inc’s MAPK genomic DNA (gDNA) reference standards represent biologically-relevant controls that can be directly incorporated into your sample processing workflows in order to optimize your protocols, evaluate assay sensitivity and specificity, and analyze the impact of workflow changes on downstream analysis. They represent ideal materials for both assay development and routine monitoring of assay performance.
The MAPK Genomic DNA Reference Standards are extracted from ASC’s panel of isogenic MAPK mutation cell lines with 50 recurrent pathway-activating mutations in the EGFR, KRAS and BRAF genes, based on data from the Sanger Institute’s COSMIC database.
Key Features of the MAPK Series gDNA Reference Standards:
Most comprehensive MAPK mutation panel on the market
Well-characterized colorectal cancer cells lines: EGFR (RKO), KRAS (RKO), BRAF (HCT116)
Paired, isogenic wild-type cell lines to serve as an ideal control
Footprint-free, homozygous mutations
Reference cell lines are expanded from single-cells, ensuring maximum homogeneity
Available in multiple formats, including slides, scrolls, and full FFPE blocks
Similar to Genetic Repair of Retinitis Pigmentosa in Patient Derived Stem Cells (20)
(June 12, 2024) Webinar: Development of PET theranostics targeting the molecu...Scintica Instrumentation
Targeting Hsp90 and its pathogen Orthologs with Tethered Inhibitors as a Diagnostic and Therapeutic Strategy for cancer and infectious diseases with Dr. Timothy Haystead.
Sexuality - Issues, Attitude and Behaviour - Applied Social Psychology - Psyc...PsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
Immersive Learning That Works: Research Grounding and Paths ForwardLeonel Morgado
We will metaverse into the essence of immersive learning, into its three dimensions and conceptual models. This approach encompasses elements from teaching methodologies to social involvement, through organizational concerns and technologies. Challenging the perception of learning as knowledge transfer, we introduce a 'Uses, Practices & Strategies' model operationalized by the 'Immersive Learning Brain' and ‘Immersion Cube’ frameworks. This approach offers a comprehensive guide through the intricacies of immersive educational experiences and spotlighting research frontiers, along the immersion dimensions of system, narrative, and agency. Our discourse extends to stakeholders beyond the academic sphere, addressing the interests of technologists, instructional designers, and policymakers. We span various contexts, from formal education to organizational transformation to the new horizon of an AI-pervasive society. This keynote aims to unite the iLRN community in a collaborative journey towards a future where immersive learning research and practice coalesce, paving the way for innovative educational research and practice landscapes.
The technology uses reclaimed CO₂ as the dyeing medium in a closed loop process. When pressurized, CO₂ becomes supercritical (SC-CO₂). In this state CO₂ has a very high solvent power, allowing the dye to dissolve easily.
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...Leonel Morgado
Current descriptions of immersive learning cases are often difficult or impossible to compare. This is due to a myriad of different options on what details to include, which aspects are relevant, and on the descriptive approaches employed. Also, these aspects often combine very specific details with more general guidelines or indicate intents and rationales without clarifying their implementation. In this paper we provide a method to describe immersive learning cases that is structured to enable comparisons, yet flexible enough to allow researchers and practitioners to decide which aspects to include. This method leverages a taxonomy that classifies educational aspects at three levels (uses, practices, and strategies) and then utilizes two frameworks, the Immersive Learning Brain and the Immersion Cube, to enable a structured description and interpretation of immersive learning cases. The method is then demonstrated on a published immersive learning case on training for wind turbine maintenance using virtual reality. Applying the method results in a structured artifact, the Immersive Learning Case Sheet, that tags the case with its proximal uses, practices, and strategies, and refines the free text case description to ensure that matching details are included. This contribution is thus a case description method in support of future comparative research of immersive learning cases. We then discuss how the resulting description and interpretation can be leveraged to change immersion learning cases, by enriching them (considering low-effort changes or additions) or innovating (exploring more challenging avenues of transformation). The method holds significant promise to support better-grounded research in immersive learning.
The cost of acquiring information by natural selectionCarl Bergstrom
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
Travis Hills of MN is Making Clean Water Accessible to All Through High Flux ...Travis Hills MN
By harnessing the power of High Flux Vacuum Membrane Distillation, Travis Hills from MN envisions a future where clean and safe drinking water is accessible to all, regardless of geographical location or economic status.
The binding of cosmological structures by massless topological defectsSérgio Sacani
Assuming spherical symmetry and weak field, it is shown that if one solves the Poisson equation or the Einstein field
equations sourced by a topological defect, i.e. a singularity of a very specific form, the result is a localized gravitational
field capable of driving flat rotation (i.e. Keplerian circular orbits at a constant speed for all radii) of test masses on a thin
spherical shell without any underlying mass. Moreover, a large-scale structure which exploits this solution by assembling
concentrically a number of such topological defects can establish a flat stellar or galactic rotation curve, and can also deflect
light in the same manner as an equipotential (isothermal) sphere. Thus, the need for dark matter or modified gravity theory is
mitigated, at least in part.
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...Advanced-Concepts-Team
Presentation in the Science Coffee of the Advanced Concepts Team of the European Space Agency on the 07.06.2024.
Speaker: Diego Blas (IFAE/ICREA)
Title: Gravitational wave detection with orbital motion of Moon and artificial
Abstract:
In this talk I will describe some recent ideas to find gravitational waves from supermassive black holes or of primordial origin by studying their secular effect on the orbital motion of the Moon or satellites that are laser ranged.
Current Ms word generated power point presentation covers major details about the micronuclei test. It's significance and assays to conduct it. It is used to detect the micronuclei formation inside the cells of nearly every multicellular organism. It's formation takes place during chromosomal sepration at metaphase.
3. Introduction
• Retinitis pigmentosa (RP)
• Inherited ( X-linked ), degenerative eye disease
• causes severe vision impairment
• progressive degeneration of the rod
photoreceptor cells in the retina
4. • The disease caused of point mutation
• C.3070 (T) at ORF15 of Retinis Rigmentosa
GTPase Regulator gene
• Stop codon stop protein synthesis
• TAG - - - - - < GAG
Stop codon- - - < Glutamate
5. Materials and Methods
• 2 technologies used in this research paper
• Genetic editing tool :
CRISPR Cas-9 ** system and
• Stem cells
Induced pluripotent stem cells
** clustered regulatory-interspaced short palindromic repeats
6. Making stem cells
• Skin-punch biopsy taken (own patient )
• Fibroblast cells taken.. Generation ability to
connective tissue
• Transcription factors added
• Induced pluripotent stem cells form (iPSCs)
• But.. iPSCs still have point mutation (TAG)
12. CRISPR Cas9
• Composed of 2 RNA guides + Cas9 endonuclease
• Transfection of iPSCs with expression vector Cas9
• Efficiency of correct cuts was 23%
• Homology-directed gene repair (HDR) completed
sequence after double strand break (DSB)
• Correction percent of the mutation (G>T) of 223
cells transfected with Cas9 was 13%
24. Conclusion
• The efficiency of cleavage of target site 3070 G>T for
CRISPR Cas9 – (g58 - gRNA) was 23% out of 293 cell
line
• Correction Rate of 23% those cells due to cell
replication proofreading machinery was 13%
according homology-directed gene repair (HDR)
• The next step is to convert corrected iPSCs to retinal
cells and transplant it in retina of the same patient
25. Reference
• Bassuk, Alexander G., Andrew Zheng, Yao Li, Stephen H.
Tsang, and Vinit B. Mahajan. "Precision Medicine: Genetic
Repair of Retinitis Pigmentosa in Patient-Derived Stem
Cells." Sci. Rep. Scientific Reports 6 (2016)