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.
An Introduction to Crispr Genome Editing
Crispr cas: A new tool of genome editing
CRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) are part of an adaptive defense mechanism in bacteria and archaea. Use of the CRISPR/Cas9 system for genome editing has been a major technological breakthrough, making genome modification in cells or organisms fast, more efficient, and much more robust than previous genome editing methods. Single guide RNAs (sgRNAs) or guide RNAs (gRNAs) direct and activate the Cas9 endonuclease at a specific genomic sequence. Cas9 then cleaves the target DNA, making it available for repair by the non-homologous end joining (NHEJ) system or for creating an insertion site for exogenous donor DNA by homologous recombination.
An Introduction to Crispr Genome Editing
Crispr cas: A new tool of genome editing
CRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) are part of an adaptive defense mechanism in bacteria and archaea. Use of the CRISPR/Cas9 system for genome editing has been a major technological breakthrough, making genome modification in cells or organisms fast, more efficient, and much more robust than previous genome editing methods. Single guide RNAs (sgRNAs) or guide RNAs (gRNAs) direct and activate the Cas9 endonuclease at a specific genomic sequence. Cas9 then cleaves the target DNA, making it available for repair by the non-homologous end joining (NHEJ) system or for creating an insertion site for exogenous donor DNA by homologous recombination.
CRISPR- Cas technology: a new antiviral weapon for plantsSachin Bhor
Genome editing is a type of genetic engineering in which DNA is inserted, replaced, or removed from a genome using artificially engineered nucleases, or "molecular scissors”. Identified CRISPR-associated protein 9 from Streptococcus pyogenes (Cas9) in (2012). Genome engineering by CRISPR/Cas system (16 February 2012). New application of this technology to combating viral infection by destroying invading virus DNA has now become possible in plants.
a brief description on the new emerging genome editing technology CRISPR-Cas9. this technique is making its place stronger and stronger day by day. and impossible things can be possible by this technique. and some main and famous names who discovered this technique.
Crispr-Cas9 system works on the concept of bacterial defence mechanism. The idea of which was replicated in eukaryotic cell in in- vitro condition by the researchers.
CRISPR- Cas technology: a new antiviral weapon for plantsSachin Bhor
Genome editing is a type of genetic engineering in which DNA is inserted, replaced, or removed from a genome using artificially engineered nucleases, or "molecular scissors”. Identified CRISPR-associated protein 9 from Streptococcus pyogenes (Cas9) in (2012). Genome engineering by CRISPR/Cas system (16 February 2012). New application of this technology to combating viral infection by destroying invading virus DNA has now become possible in plants.
a brief description on the new emerging genome editing technology CRISPR-Cas9. this technique is making its place stronger and stronger day by day. and impossible things can be possible by this technique. and some main and famous names who discovered this technique.
Crispr-Cas9 system works on the concept of bacterial defence mechanism. The idea of which was replicated in eukaryotic cell in in- vitro condition by the researchers.
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)Akshay Deshmukh
clustered regularly interspaced short palindromic repeats is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria. Now CRISPR use as genome editing tool in different Plant Breeder to manipulate the DNA of the crop
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
How CRISPR–Cas9 Screening will revolutionise your drug development programsHorizonDiscovery
CRISPR–Cas9 mediated genome editing provides a novel and 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. This makes CRISPR–Cas9 screening a powerful tool for drug target ID and validation, understanding drug mechanisms of action and patient stratification.
In this webinar, we use our experience with CRISPR–Cas9 to discuss the power and applicability of CRISPR-Cas9 screening technologies. We focus on how to use this technology to address important biological questions, and consider what’s possible, what’s plausible and what constitutes a ‘hit’. We also highlight Horizon’s latest developments to the CRISPR-Cas9 screening platform.
Codons Optimization by Creative BiogeneDonglin Bao
Codon optimization is one of the key steps in achieving the high-level expression of target gene. There are some key factors for consideration including transcription efficiency, translation efficiency, gene synthesis and protein folding. Creative Biogene can optimize almost perfectly these key factors in codon optimization.
Principle and method of RNAi-Creative BiogeneDonglin Bao
In recent years, studies have shown that mRNA will occur specific degradation if dsRNA which is composed of sense RNA and antisense RNA from mRNA is transferred into the cell, eventually it leads to target genes silence. The post-transcriptional gene silencing (PTGS) is described as RNAi.
ATAC-seq (Assay for Transposase-Accessible Chromatin using sequencing) is a technique used in molecular biology to study chromatin accessibility. The key part of the ATAC-seq procedure is the action of the transposase Tn5 on the genomic DNA of the sample.
Codon optimization is one of the key steps in achieving the high level expression of target gene. There are some key factors for consideration including transcription efficiency, translation efficiency, gene synthesis and protein folding.
RT-PCR (reverse transcription-polymerase chain reaction) is a variant of the polymerase chain reaction (PCR) which are now widely used. Traditionally RT-PCR involves two steps: the RT reaction and PCR amplification. RNA is first reverse transcribed into cDNA using a reverse transcriptase as described here, the resulting cDNA is used as templates for subsequent PCR amplification using primers specific for one or more genes. RT-PCR can be used to quantify mRNA levels from much smaller samples. In fact, this technique is sensitive enough to enable quantitation of RNA from a single cell.
Gene overexpression is the process which leads to the abundant target protein expression subsequently. The process may be in the cell where the gene is originally located or in other expression systems. The fundamental principle is to add regulatory elements to the upstream of the target gene through artificial construction, so that genes can be transcribed and translated efficiently under controlled conditions.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
2. Three Generation of Gene editing tools
Liu, J. et al, 2015, Nat Protoc
Three Generation of Gene editing tools:
Zinc Finger Nuclease (ZFN)
Transcription Activator Like Effector Nuclease (TALEN)
Clustered Regularly Interspaced Short Palindromic Repeats associated systems (CRISPR/Cas9)
3. CRISPR: The scientific breakthrough of 2015
Adapted from Feng Zhang et al, 2015, Cell
The applications of Gene editing tools:
CRISPR makes gene editing easily and
effectively, no matter in Biological Research or
technological application.
4. What is CRISPR?
The mechanism of CRISPR/Cas9 system:
Ran et al, 2014, Nat Proto
Cas9 could cleave the target site and cause a Double Stranded Break(DSB).
5. Non-Homologous End Joining (NHEJ)
Homologous Directed Repair (HDR)
DNA repair mechanism
Two main mechanisms for repairing DSB
Gene Knockout
Gene replacement/Knockin
Without homologous template
With homologous template
Ran et al, 2014, Nat Proto
6. Main applications of CRISPR/Cas9 technology
Gene KO
Reporter Gene KI
Gene KI in Mouse Embryos
Gene KI in Stem cells
Gene Deletion
7. Example：CFTR editing by using CRISPR/Cas9
Correction of F508del by CRISPR-induced HDR
Schwank et.al, 2013, Cell Stem Cell
Cystic Fibrosis is a genetic disorder
caused by gene mutations in CFTR gene,
especially F508 deletion.
8. Procedure: Let CRISPR help your research
Adapted from Swati Kadam, 2016, ThermoFisher Scientific Website
Visit our website: Creative Biogene for more resources and protocols
