mRNA Processing in Eukaryotes
Spliceosom Activity
Alternative Splicing
Self Splicing
Post Transcriptional Modifications
Prep by Shahzaib Khurshid
BS ZOOLOGY
Punjab University Lahore
Pyrosequencing slide presentation rev3.Robert Bruce
Pyrosequencing was evaluated as an alternative to Sanger sequencing for HIV drug resistance genotyping. Three assays were designed to detect mutations in the protease gene using a 356 bp amplicon and three sequencing primers. The assays demonstrated good linearity and sensitivity below 5% in quantifying mixed bases. However, read lengths were limited due to signal degradation, making it difficult to sequence large amplicons. Overall, pyrosequencing showed promise as a method for HIV genotyping but further optimization was needed to improve read lengths and mixed variant detection.
This document discusses different types of genetic recombination enzymes called recombinases. It describes two main types of recombination - homologous recombination, which involves exchange between similar DNA sequences, and site-specific recombination, which recombines DNA at specific sites regardless of homology. The key recombinases involved in homologous recombination are RecBCD and RecA in E. coli. There are two families of site-specific recombinases - serine recombinases like Hin that cleave all DNA strands, and tyrosine recombinases like Cre, Lambda and Xer that cleave strands one pair at a time.
The pentose phosphate pathway generates NADPH and pentose sugars. Glucose-6-phosphate enters the pathway and is oxidized by glucose-6-phosphate dehydrogenase, producing 6-phosphogluconolactone. This is hydrolyzed to 6-phosphogluconate by 6-phosphogluconolactonase. 6-Phosphogluconate dehydrogenase further oxidizes this to ribulose-5-phosphate, producing NADPH. Ribulose-5-phosphate can be converted to other 5-carbon sugars like ribose-5-phosphate. Transketolase and transaldolase reactions interconvert pentose and triose phosphates to regenerate ribulose
Accurate DNA Methylation Analysis with Successful Bisulfite Conversion WebinarQIAGEN
Bisulfite conversion is a popular used method for DNA methylation analysis. It is the most convenient and effective way to map DNA methylation to individual bases. The efficiency of the bisulfite conversion has a huge impact on the reliability of the downstream analysis methods and complete conversion is a prerequisite for correct determination of methylation. However, standard methods require harsh conditions with long incubations times at high temperatures and low pH to achieve complete conversion. This harsh treatment can lead to DNA degradation, lowering the yields and sensitivity of the subsequent analysis.
This slidedeck:
• Explains the principle of bisulfite conversion
• Points out the challenges and critical factors for successful bisulfite conversion
• Describes how to overcome the challenges with QIAGEN’s EpiTect Fast Bisulfite Kits
• Gives general recommendations for planning successful bisulfite conversion experiments
The document discusses gene regulation and provides examples in bacteria. It explains that certain genes are turned on or off depending on environmental factors. It describes the lac operon in E. coli, which contains genes involved in processing the sugar lactose. The lac operon is regulated by the presence of lactose and glucose such that the genes are only expressed when glucose is absent and lactose is present.
The document discusses various types of DNA damage including deamination, depurination, UV light-induced T-T and T-C dimers, alkylation, oxidative damage, replication errors, and double-strand breaks. It then summarizes different DNA repair pathways such as base excision repair, nucleotide excision repair, mismatch repair, direct repair, recombination repair, and non-homologous end-joining. The SOS response in bacteria is also summarized as activating error-prone repair when normal repair pathways are overwhelmed.
This document discusses two approaches to pyrosequencing technology - solid phase and liquid phase. The solid phase approach utilizes streptavidin coated beads to immobilize biotin-labeled DNA templates. It involves sequential addition of nucleotides followed by washing steps to remove unincoporated nucleotides. The liquid phase approach introduced an enzyme called apyrase that degrades unincorporated nucleotides, eliminating the need for washing steps. It involves a cascade of four enzymes - DNA polymerase, ATP sulfurylase, luciferase, and apyrase to continuously degrade unincorporated nucleotides and determine DNA sequences from light signals.
Pyrosequencing slide presentation rev3.Robert Bruce
Pyrosequencing was evaluated as an alternative to Sanger sequencing for HIV drug resistance genotyping. Three assays were designed to detect mutations in the protease gene using a 356 bp amplicon and three sequencing primers. The assays demonstrated good linearity and sensitivity below 5% in quantifying mixed bases. However, read lengths were limited due to signal degradation, making it difficult to sequence large amplicons. Overall, pyrosequencing showed promise as a method for HIV genotyping but further optimization was needed to improve read lengths and mixed variant detection.
This document discusses different types of genetic recombination enzymes called recombinases. It describes two main types of recombination - homologous recombination, which involves exchange between similar DNA sequences, and site-specific recombination, which recombines DNA at specific sites regardless of homology. The key recombinases involved in homologous recombination are RecBCD and RecA in E. coli. There are two families of site-specific recombinases - serine recombinases like Hin that cleave all DNA strands, and tyrosine recombinases like Cre, Lambda and Xer that cleave strands one pair at a time.
The pentose phosphate pathway generates NADPH and pentose sugars. Glucose-6-phosphate enters the pathway and is oxidized by glucose-6-phosphate dehydrogenase, producing 6-phosphogluconolactone. This is hydrolyzed to 6-phosphogluconate by 6-phosphogluconolactonase. 6-Phosphogluconate dehydrogenase further oxidizes this to ribulose-5-phosphate, producing NADPH. Ribulose-5-phosphate can be converted to other 5-carbon sugars like ribose-5-phosphate. Transketolase and transaldolase reactions interconvert pentose and triose phosphates to regenerate ribulose
Accurate DNA Methylation Analysis with Successful Bisulfite Conversion WebinarQIAGEN
Bisulfite conversion is a popular used method for DNA methylation analysis. It is the most convenient and effective way to map DNA methylation to individual bases. The efficiency of the bisulfite conversion has a huge impact on the reliability of the downstream analysis methods and complete conversion is a prerequisite for correct determination of methylation. However, standard methods require harsh conditions with long incubations times at high temperatures and low pH to achieve complete conversion. This harsh treatment can lead to DNA degradation, lowering the yields and sensitivity of the subsequent analysis.
This slidedeck:
• Explains the principle of bisulfite conversion
• Points out the challenges and critical factors for successful bisulfite conversion
• Describes how to overcome the challenges with QIAGEN’s EpiTect Fast Bisulfite Kits
• Gives general recommendations for planning successful bisulfite conversion experiments
The document discusses gene regulation and provides examples in bacteria. It explains that certain genes are turned on or off depending on environmental factors. It describes the lac operon in E. coli, which contains genes involved in processing the sugar lactose. The lac operon is regulated by the presence of lactose and glucose such that the genes are only expressed when glucose is absent and lactose is present.
The document discusses various types of DNA damage including deamination, depurination, UV light-induced T-T and T-C dimers, alkylation, oxidative damage, replication errors, and double-strand breaks. It then summarizes different DNA repair pathways such as base excision repair, nucleotide excision repair, mismatch repair, direct repair, recombination repair, and non-homologous end-joining. The SOS response in bacteria is also summarized as activating error-prone repair when normal repair pathways are overwhelmed.
This document discusses two approaches to pyrosequencing technology - solid phase and liquid phase. The solid phase approach utilizes streptavidin coated beads to immobilize biotin-labeled DNA templates. It involves sequential addition of nucleotides followed by washing steps to remove unincoporated nucleotides. The liquid phase approach introduced an enzyme called apyrase that degrades unincorporated nucleotides, eliminating the need for washing steps. It involves a cascade of four enzymes - DNA polymerase, ATP sulfurylase, luciferase, and apyrase to continuously degrade unincorporated nucleotides and determine DNA sequences from light signals.
Colony hybridization is a technique to identify bacterial colonies containing a specific DNA sequence or gene of interest. It involves transferring DNA from bacterial colonies onto a membrane, then probing the membrane with a complementary DNA or RNA sequence. Only colonies with matching DNA sequences will hybridize with the probe. The oligonucleotide ligation assay (OLA) is a technique used to detect mutations by hybridizing PCR primers and ligating adjacent probes only when the target sequence is present. It has advantages of being rapid, easy, and high-throughput but requires an automated sequencer.
Role of Antisense and RNAi-based Gene Silencing in Crop ImprovementMariya Zaman
This document presents information on RNA interference (RNAi) and its application in crop improvement. It discusses the discovery of antisense RNA and RNAi technology. The mechanisms of antisense technology and RNAi are described. Advantages of RNAi include its ability to study essential genes and its high specificity. Applications include crop protection and gene therapy. Case studies demonstrate improved insect resistance in transgenic tobacco plants and the role of miRNAs in syncytium formation induced by cyst nematodes.
Applications and drawbacks of sanger sequencingHome
Sanger sequencing is a method for determining the order of bases in DNA developed by Fred Sanger in 1977. It involves making copies of a DNA region using DNA polymerase along with regular and chain-terminating dideoxynucleotides labeled with different dyes. This allows sequences of up to 900 base pairs to be determined. Sanger sequencing has applications in SNP detection, SSCP analysis, and STR analysis but is limited by only being able to sequence short DNA fragments and being relatively expensive compared to newer sequencing methods.
The document summarizes the Ramachandran plot, which is a plot of the phi and psi dihedral angles of amino acid residues in protein structures. It was originally developed in 1963 to show possible conformations of phi and psi angles for amino acid residues and the empirical distribution of these angles observed in protein structures. The plot takes advantage of the circular nature of dihedral angles, with edges wrapping from right to left and bottom to top. It can be used to determine amino acid preferences and how the presence or absence of groups like a methylene group at C-beta affect the angles.
The document summarizes transcription in prokaryotes. It discusses the key components including the template strand, coding strand, and RNA polymerase. RNA polymerase is made up of multiple subunits and recognizes promoter sequences to initiate transcription. The process of transcription involves three phases - initiation when RNA polymerase binds to the promoter, elongation as the RNA strand continuously grows, and termination when RNA polymerase stops synthesis.
The pyruvate dehydrogenase complex (PDC) catalyzes the oxidative decarboxylation of pyruvate to form acetyl-CoA and consists of multiple enzymes and cofactors. Pyruvate dehydrogenase (E1) uses thiamine pyrophosphate to decarboxylate pyruvate. Dihydrolipoyl transacetylase (E2) transfers the acetyl group to coenzyme A with help from lipoic acid. Dihydrolipoyl dehydrogenase (E3) regenerates the oxidized cofactors using NAD+ and FAD, generating NADH to fuel the electron transport chain. PDC is regulated by product inhibition and phosphorylation/dephosphorylation of E1 by
The present ppt is covers all aspects of protein translation in bacteria as well as in eukaryotes. It also includes a brief introduction to ribosomes and tRNA which are among the key components of the translation machinery.
1) DNA replication begins with the unwinding of the DNA double helix at an origin of replication site.
2) This forms a replication fork with leading and lagging strands that are copied semi-conservatively to produce two identical copies of DNA.
3) RNA primers, DNA polymerases, helicase and single-strand binding proteins work together to separate the strands and synthesize new DNA in the 5’-3’ direction along the template.
Translation is the process by which the genetic code in mRNA is used to direct the synthesis of proteins. It involves three main steps - initiation, elongation, and termination. Initiation requires the small and large ribosomal subunits to assemble around an mRNA molecule along with initiator tRNA and other initiation factors. Elongation then adds amino acids one by one to the growing polypeptide chain according to the mRNA codons. Termination occurs when a stop codon is reached, causing the ribosome to dissociate and release the complete protein.
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.
In situ hybridization is a powerful technique for identifying specific mRNA species within individual cells in tissue sections, providing insights into physiological processes and disease pathogenesis. However, in situ hybridization requires that many steps be taken with precise optimization for each tissue examined and for each probe used.
https://www.creative-bioarray.com/Services/In-Situ-Hybridization-ISH.htm
This document summarizes a presentation on the role of antisense and RNA interference (RNAi) gene silencing in crop improvement. It discusses the discovery of RNAi as a mechanism of gene silencing triggered by double-stranded RNA, and how this natural process can be harnessed through genetic engineering to modify traits in crops. Examples are given of crops where RNAi has been used to develop tolerance to herbicides, modify starch composition, or reduce virus susceptibility. The document concludes that transgenic RNA silencing avoids risks associated with foreign proteins and can be easily introduced into hybrid crops.
This document discusses RNA editing. It defines RNA editing as molecular processes that alter the nucleotide sequence of an RNA molecule after transcription from DNA. There are two main types of RNA editing: substitution editing, where individual nucleotides are chemically altered, and insertion/deletion editing, which adds or removes nucleotides. An example of substitution editing is provided in the human APOB gene, where an enzyme changes a codon resulting in alternative protein isoforms. Guide RNAs are also discussed as facilitating insertion/deletion editing through base-pairing with pre-mRNAs. RNA editing increases proteomic diversity and regulates gene expression.
Pyrosequencing is a sequencing by synthesis technique that uses a luciferase enzyme system to monitor DNA synthesis. It works by adding DNA polymerase and a single nucleotide to the DNA fragments, generating pyrophosphate that is converted to light. The light is detected and identifies the nucleotide incorporated. Pyrosequencing has applications in cDNA analysis, mutation detection, re-sequencing of disease genes, and identifying single nucleotide polymorphisms and typing bacteria and viruses.
DNA replication is the process by which a cell makes an identical copy of its DNA. It occurs during the S phase of the cell cycle and involves unwinding the DNA double helix, creating RNA primers, and synthesizing new DNA strands using existing strands as templates in the 5' to 3' direction. The replication factory contains many replication proteins that cluster together to duplicate DNA. Experimental evidence from Meselson-Stahl experiments supported the semi-conservative mode of replication, in which each new DNA molecule contains one original and one new strand. Replication initiates at specific origins of replication and proceeds bidirectionally.
This document provides an overview of antisense technology for inhibiting gene expression. It discusses how antisense oligonucleotides are complementary to mRNA and can bind to prevent translation. The mechanisms of hybridization arrest and RNaseH activation are described. Different approaches using antisense oligonucleotides that do and do not activate RNaseH are covered. Applications in agriculture and medicine are mentioned, including the development of the Flavr Savr tomato that used antisense technology to delay fruit ripening. The role of antisense oligonucleotides in drug discovery is also briefly discussed.
The document discusses various processes involved in eukaryotic mRNA processing. It explains that in eukaryotes, pre-mRNA undergoes 5' capping, 3' cleavage and polyadenylation, splicing, and methylation to become a mature mRNA. These processes are guided by hnRNP and snRNP particles. The document also discusses alternative mRNA processing mechanisms like alternative splicing and polyadenylation that generate multiple mRNA isoforms from a single pre-mRNA.
This document summarizes the process of gene expression from DNA to protein. It discusses:
1) How early experiments established that genes encode proteins through RNA intermediates. One gene encodes one polypeptide.
2) The "central dogma" of molecular biology - that DNA is transcribed into RNA which is then translated into protein. RNA acts as an intermediate to protect DNA and allow for gene regulation.
3) The process of transcription, including initiation, elongation, termination, and post-transcriptional modification of pre-mRNA in eukaryotes.
4) Translation occurs on ribosomes, where mRNA directs the assembly of amino acids into polypeptides according to the genetic code
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.
Colony hybridization is a technique to identify bacterial colonies containing a specific DNA sequence or gene of interest. It involves transferring DNA from bacterial colonies onto a membrane, then probing the membrane with a complementary DNA or RNA sequence. Only colonies with matching DNA sequences will hybridize with the probe. The oligonucleotide ligation assay (OLA) is a technique used to detect mutations by hybridizing PCR primers and ligating adjacent probes only when the target sequence is present. It has advantages of being rapid, easy, and high-throughput but requires an automated sequencer.
Role of Antisense and RNAi-based Gene Silencing in Crop ImprovementMariya Zaman
This document presents information on RNA interference (RNAi) and its application in crop improvement. It discusses the discovery of antisense RNA and RNAi technology. The mechanisms of antisense technology and RNAi are described. Advantages of RNAi include its ability to study essential genes and its high specificity. Applications include crop protection and gene therapy. Case studies demonstrate improved insect resistance in transgenic tobacco plants and the role of miRNAs in syncytium formation induced by cyst nematodes.
Applications and drawbacks of sanger sequencingHome
Sanger sequencing is a method for determining the order of bases in DNA developed by Fred Sanger in 1977. It involves making copies of a DNA region using DNA polymerase along with regular and chain-terminating dideoxynucleotides labeled with different dyes. This allows sequences of up to 900 base pairs to be determined. Sanger sequencing has applications in SNP detection, SSCP analysis, and STR analysis but is limited by only being able to sequence short DNA fragments and being relatively expensive compared to newer sequencing methods.
The document summarizes the Ramachandran plot, which is a plot of the phi and psi dihedral angles of amino acid residues in protein structures. It was originally developed in 1963 to show possible conformations of phi and psi angles for amino acid residues and the empirical distribution of these angles observed in protein structures. The plot takes advantage of the circular nature of dihedral angles, with edges wrapping from right to left and bottom to top. It can be used to determine amino acid preferences and how the presence or absence of groups like a methylene group at C-beta affect the angles.
The document summarizes transcription in prokaryotes. It discusses the key components including the template strand, coding strand, and RNA polymerase. RNA polymerase is made up of multiple subunits and recognizes promoter sequences to initiate transcription. The process of transcription involves three phases - initiation when RNA polymerase binds to the promoter, elongation as the RNA strand continuously grows, and termination when RNA polymerase stops synthesis.
The pyruvate dehydrogenase complex (PDC) catalyzes the oxidative decarboxylation of pyruvate to form acetyl-CoA and consists of multiple enzymes and cofactors. Pyruvate dehydrogenase (E1) uses thiamine pyrophosphate to decarboxylate pyruvate. Dihydrolipoyl transacetylase (E2) transfers the acetyl group to coenzyme A with help from lipoic acid. Dihydrolipoyl dehydrogenase (E3) regenerates the oxidized cofactors using NAD+ and FAD, generating NADH to fuel the electron transport chain. PDC is regulated by product inhibition and phosphorylation/dephosphorylation of E1 by
The present ppt is covers all aspects of protein translation in bacteria as well as in eukaryotes. It also includes a brief introduction to ribosomes and tRNA which are among the key components of the translation machinery.
1) DNA replication begins with the unwinding of the DNA double helix at an origin of replication site.
2) This forms a replication fork with leading and lagging strands that are copied semi-conservatively to produce two identical copies of DNA.
3) RNA primers, DNA polymerases, helicase and single-strand binding proteins work together to separate the strands and synthesize new DNA in the 5’-3’ direction along the template.
Translation is the process by which the genetic code in mRNA is used to direct the synthesis of proteins. It involves three main steps - initiation, elongation, and termination. Initiation requires the small and large ribosomal subunits to assemble around an mRNA molecule along with initiator tRNA and other initiation factors. Elongation then adds amino acids one by one to the growing polypeptide chain according to the mRNA codons. Termination occurs when a stop codon is reached, causing the ribosome to dissociate and release the complete protein.
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.
In situ hybridization is a powerful technique for identifying specific mRNA species within individual cells in tissue sections, providing insights into physiological processes and disease pathogenesis. However, in situ hybridization requires that many steps be taken with precise optimization for each tissue examined and for each probe used.
https://www.creative-bioarray.com/Services/In-Situ-Hybridization-ISH.htm
This document summarizes a presentation on the role of antisense and RNA interference (RNAi) gene silencing in crop improvement. It discusses the discovery of RNAi as a mechanism of gene silencing triggered by double-stranded RNA, and how this natural process can be harnessed through genetic engineering to modify traits in crops. Examples are given of crops where RNAi has been used to develop tolerance to herbicides, modify starch composition, or reduce virus susceptibility. The document concludes that transgenic RNA silencing avoids risks associated with foreign proteins and can be easily introduced into hybrid crops.
This document discusses RNA editing. It defines RNA editing as molecular processes that alter the nucleotide sequence of an RNA molecule after transcription from DNA. There are two main types of RNA editing: substitution editing, where individual nucleotides are chemically altered, and insertion/deletion editing, which adds or removes nucleotides. An example of substitution editing is provided in the human APOB gene, where an enzyme changes a codon resulting in alternative protein isoforms. Guide RNAs are also discussed as facilitating insertion/deletion editing through base-pairing with pre-mRNAs. RNA editing increases proteomic diversity and regulates gene expression.
Pyrosequencing is a sequencing by synthesis technique that uses a luciferase enzyme system to monitor DNA synthesis. It works by adding DNA polymerase and a single nucleotide to the DNA fragments, generating pyrophosphate that is converted to light. The light is detected and identifies the nucleotide incorporated. Pyrosequencing has applications in cDNA analysis, mutation detection, re-sequencing of disease genes, and identifying single nucleotide polymorphisms and typing bacteria and viruses.
DNA replication is the process by which a cell makes an identical copy of its DNA. It occurs during the S phase of the cell cycle and involves unwinding the DNA double helix, creating RNA primers, and synthesizing new DNA strands using existing strands as templates in the 5' to 3' direction. The replication factory contains many replication proteins that cluster together to duplicate DNA. Experimental evidence from Meselson-Stahl experiments supported the semi-conservative mode of replication, in which each new DNA molecule contains one original and one new strand. Replication initiates at specific origins of replication and proceeds bidirectionally.
This document provides an overview of antisense technology for inhibiting gene expression. It discusses how antisense oligonucleotides are complementary to mRNA and can bind to prevent translation. The mechanisms of hybridization arrest and RNaseH activation are described. Different approaches using antisense oligonucleotides that do and do not activate RNaseH are covered. Applications in agriculture and medicine are mentioned, including the development of the Flavr Savr tomato that used antisense technology to delay fruit ripening. The role of antisense oligonucleotides in drug discovery is also briefly discussed.
The document discusses various processes involved in eukaryotic mRNA processing. It explains that in eukaryotes, pre-mRNA undergoes 5' capping, 3' cleavage and polyadenylation, splicing, and methylation to become a mature mRNA. These processes are guided by hnRNP and snRNP particles. The document also discusses alternative mRNA processing mechanisms like alternative splicing and polyadenylation that generate multiple mRNA isoforms from a single pre-mRNA.
This document summarizes the process of gene expression from DNA to protein. It discusses:
1) How early experiments established that genes encode proteins through RNA intermediates. One gene encodes one polypeptide.
2) The "central dogma" of molecular biology - that DNA is transcribed into RNA which is then translated into protein. RNA acts as an intermediate to protect DNA and allow for gene regulation.
3) The process of transcription, including initiation, elongation, termination, and post-transcriptional modification of pre-mRNA in eukaryotes.
4) Translation occurs on ribosomes, where mRNA directs the assembly of amino acids into polypeptides according to the genetic code
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.
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!
Authoring a personal GPT for your research and practice: How we created the Q...Leonel Morgado
Thematic analysis in qualitative research is a time-consuming and systematic task, typically done using teams. Team members must ground their activities on common understandings of the major concepts underlying the thematic analysis, and define criteria for its development. However, conceptual misunderstandings, equivocations, and lack of adherence to criteria are challenges to the quality and speed of this process. Given the distributed and uncertain nature of this process, we wondered if the tasks in thematic analysis could be supported by readily available artificial intelligence chatbots. Our early efforts point to potential benefits: not just saving time in the coding process but better adherence to criteria and grounding, by increasing triangulation between humans and artificial intelligence. This tutorial will provide a description and demonstration of the process we followed, as two academic researchers, to develop a custom ChatGPT to assist with qualitative coding in the thematic data analysis process of immersive learning accounts in a survey of the academic literature: QUAL-E Immersive Learning Thematic Analysis Helper. In the hands-on time, participants will try out QUAL-E and develop their ideas for their own qualitative coding ChatGPT. Participants that have the paid ChatGPT Plus subscription can create a draft of their assistants. The organizers will provide course materials and slide deck that participants will be able to utilize to continue development of their custom GPT. The paid subscription to ChatGPT Plus is not required to participate in this workshop, just for trying out personal GPTs during it.
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.
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
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.
Mending Clothing to Support Sustainable Fashion_CIMaR 2024.pdfSelcen Ozturkcan
Ozturkcan, S., Berndt, A., & Angelakis, A. (2024). Mending clothing to support sustainable fashion. Presented at the 31st Annual Conference by the Consortium for International Marketing Research (CIMaR), 10-13 Jun 2024, University of Gävle, Sweden.
PPT on Direct Seeded Rice presented at the three-day 'Training and Validation Workshop on Modules of Climate Smart Agriculture (CSA) Technologies in South Asia' workshop on April 22, 2024.
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.
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.
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