It is very fast and new technique for detection and degradation of viral DNA and it is so helpful for us to understand how to degraded viral DNA... what type of function naturally present in bacteria........ so its very excellent technique
CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found within the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that have previously infected the prokaryote and are used to detect and destroy DNA from similar phages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.
Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within organisms.This editing process has a wide variety of applications including basic biological research, development of biotechnology products, and treatment of diseases.
The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.
This document discusses the CRISPR-Cas9 genome editing technique. It begins with an introduction to CRISPR as an adaptive immune system in bacteria. The CRISPR mechanism involves acquiring DNA from invading viruses and using CRISPR RNA and Cas9 proteins to cut matching viral DNA. Scientists now use the Cas9 nuclease guided by a synthetic single guide RNA to make targeted cuts in DNA for genetic engineering. Some applications include modifying crop plants and research in mice embryos. However, using CRISPR in human embryos raises ethical concerns about germline editing and unintended consequences.
The document discusses genome sequencing and related topics. It begins by defining what a genome is - the complete set of DNA in an organism. It then discusses the different types of genomes, such as prokaryotic and eukaryotic, including nuclear, mitochondrial, and chloroplast genomes. The document also defines genomics as the comprehensive study of whole genomes and all gene interactions, distinguishing it from traditional genetics which focuses on single genes. It outlines some key milestones in genomic sequencing and the technical foundations that enabled sequencing whole genomes. Finally, it describes the main approaches used for genome sequencing projects, including hierarchical shotgun sequencing and whole genome shotgun sequencing.
complete Single Nucleotide Polymorphiitsm Detection methods with Advance techniques with its applications
Single nucleotide polymorphisms are single base variations between genomes within a species.
There are at least 10 million polymorphic sites in the human genome.
SNPs can distinguish individuals from one another
Denaturing Gradient Gel Electrophoresis
Chemical Cleavage Of Mismatch
Single-stranded Conformation Polymorphism (SSCP)
MutS Protein-binding Assays
Mismatch Repair Detection (MRD)
Heteroduplex Analysis (HA)
Denaturing High Performance Liquid Chromatography (DHPLC)
UNG-Mediated T-Sequencing
RNA-Mediated Finger printing with MALDI MS Detection
Sequencing by Hybridization
Direct DNA Sequencing
Single-feature polymorphism (SFP)
Invader probe
Allele-specific oligonucleotide probes
PCR-based methods
Allele specific primers
Sequence Polymorphism-Derived (SPD) markers
Targeting induced local lesions in genomes (TILLinG)
Minisequencing primers
Allele-specific ligation probes
Genome editing with the CRISPR-Cas9 system has become one of the major tools in modern biotechnology. This slide share discusses the fundamentals in a simple, easy to understand format.
Crispr cas: A new tool of genome editing palaabhay
The document summarizes a presentation on CRISPR cas9, a new genome editing tool. It discusses the history of CRISPR, how CRISPR functions in bacteria, the classification and components of CRISPR systems, and the mechanism of CRISPR cas9. It then covers applications of CRISPR cas9 in genome editing, databases of CRISPR sequences, case studies using the technology, and future directions of CRISPR research.
Gene editing using CRISPR was originally discovered as a bacterial immune system that provides resistance to viruses. CRISPR uses specialized DNA sequences and associated Cas proteins to create targeted double-strand breaks in DNA, allowing modification of genomes. The technique has rapidly advanced due to its simplicity and versatility compared to prior tools. CRISPR holds promise for treating genetic diseases, transplantation, biotechnology, disease models, and more. It has become a widely used research tool with many companies and publications emerging around its applications.
CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA sequences found within the genomes of prokaryotic organisms such as bacteria and archaea. These sequences are derived from DNA fragments of bacteriophages that have previously infected the prokaryote and are used to detect and destroy DNA from similar phages during subsequent infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.
Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within organisms.This editing process has a wide variety of applications including basic biological research, development of biotechnology products, and treatment of diseases.
The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of sequenced archaea.
This document discusses the CRISPR-Cas9 genome editing technique. It begins with an introduction to CRISPR as an adaptive immune system in bacteria. The CRISPR mechanism involves acquiring DNA from invading viruses and using CRISPR RNA and Cas9 proteins to cut matching viral DNA. Scientists now use the Cas9 nuclease guided by a synthetic single guide RNA to make targeted cuts in DNA for genetic engineering. Some applications include modifying crop plants and research in mice embryos. However, using CRISPR in human embryos raises ethical concerns about germline editing and unintended consequences.
The document discusses genome sequencing and related topics. It begins by defining what a genome is - the complete set of DNA in an organism. It then discusses the different types of genomes, such as prokaryotic and eukaryotic, including nuclear, mitochondrial, and chloroplast genomes. The document also defines genomics as the comprehensive study of whole genomes and all gene interactions, distinguishing it from traditional genetics which focuses on single genes. It outlines some key milestones in genomic sequencing and the technical foundations that enabled sequencing whole genomes. Finally, it describes the main approaches used for genome sequencing projects, including hierarchical shotgun sequencing and whole genome shotgun sequencing.
complete Single Nucleotide Polymorphiitsm Detection methods with Advance techniques with its applications
Single nucleotide polymorphisms are single base variations between genomes within a species.
There are at least 10 million polymorphic sites in the human genome.
SNPs can distinguish individuals from one another
Denaturing Gradient Gel Electrophoresis
Chemical Cleavage Of Mismatch
Single-stranded Conformation Polymorphism (SSCP)
MutS Protein-binding Assays
Mismatch Repair Detection (MRD)
Heteroduplex Analysis (HA)
Denaturing High Performance Liquid Chromatography (DHPLC)
UNG-Mediated T-Sequencing
RNA-Mediated Finger printing with MALDI MS Detection
Sequencing by Hybridization
Direct DNA Sequencing
Single-feature polymorphism (SFP)
Invader probe
Allele-specific oligonucleotide probes
PCR-based methods
Allele specific primers
Sequence Polymorphism-Derived (SPD) markers
Targeting induced local lesions in genomes (TILLinG)
Minisequencing primers
Allele-specific ligation probes
Genome editing with the CRISPR-Cas9 system has become one of the major tools in modern biotechnology. This slide share discusses the fundamentals in a simple, easy to understand format.
Crispr cas: A new tool of genome editing palaabhay
The document summarizes a presentation on CRISPR cas9, a new genome editing tool. It discusses the history of CRISPR, how CRISPR functions in bacteria, the classification and components of CRISPR systems, and the mechanism of CRISPR cas9. It then covers applications of CRISPR cas9 in genome editing, databases of CRISPR sequences, case studies using the technology, and future directions of CRISPR research.
Gene editing using CRISPR was originally discovered as a bacterial immune system that provides resistance to viruses. CRISPR uses specialized DNA sequences and associated Cas proteins to create targeted double-strand breaks in DNA, allowing modification of genomes. The technique has rapidly advanced due to its simplicity and versatility compared to prior tools. CRISPR holds promise for treating genetic diseases, transplantation, biotechnology, disease models, and more. It has become a widely used research tool with many companies and publications emerging around its applications.
This document discusses genome editing techniques. It begins by defining genomes and how they consist of DNA or RNA that contains both coding and non-coding regions. It then discusses several methods of genome editing including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR-Cas system. Each method uses engineered nucleases to introduce targeted double-strand breaks in DNA, allowing the cell's repair mechanisms to modify the genome. The CRISPR-Cas system was selected as the breakthrough of the year in 2015 due to its simplicity, efficiency and precision for genome editing applications.
Comparative genomics involves comparing genomes to discover similarities and differences. It can provide insights into evolutionary relationships, help predict gene function, and aid in drug discovery. The first step is often aligning genome sequences using tools like BLAST or MUMmer. Genomes can then be compared at various levels, such as overall nucleotide statistics, genome structure, and coding/non-coding regions. Comparing gene and protein content across genomes helps predict functions. Conserved genomic features across species also aid prediction. Insights into genome evolution come from studying molecular events like inversions and duplications. Comparative genomics has impacted phylogenetics and drug target identification.
Genome editing techniques allow DNA to be inserted, deleted, modified or replaced in the genome of a living organism. Four families of engineered nucleases have been used for genome editing: meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR/Cas9 system. Each system uses a different mechanism for recognizing and cutting DNA at specific locations to modify genes. The CRISPR/Cas9 system has become widely used due to its ease of design and lower off-target effects compared to other techniques.
This document discusses forward and reverse genetic approaches for understanding gene function. Forward genetics begins with a phenotype and identifies the underlying gene, while reverse genetics starts with a gene and determines its phenotype. Specific reverse genetic techniques described include large-scale random mutagenesis, homologous recombination, transposable element excision, RNA interference, genome editing using ZFNs/TALENs/CRISPR, and site-directed mutagenesis combined with transgenics. The document provides details on how each technique is used to alter genes and study their function.
Zinc finger nucleases (ZFNs) allow for highly targeted editing of the genome. ZFNs consist of a DNA-binding domain made of zinc finger proteins and a DNA-cleaving domain. The ZFN pair binds to a target site and creates a double-strand break, which the cell repairs through non-homologous end joining or homologous recombination, enabling gene knockouts or targeted changes. ZFNs work in many cell types and animal models, providing a more efficient alternative to traditional transgenic techniques. They have applications in functional genomics, cell line engineering, and animal model generation.
ESTs are short sequences of DNA derived from cDNA clones that represent gene expression in particular cells or tissues. They provide a simple and inexpensive way to discover new genes and map their positions in genomes. To create an EST, mRNA is converted to cDNA and then sequenced, yielding short expressed DNA sequences. ESTs are deposited in public databases like NCBI's dbEST and can help identify genes, construct genome maps, and characterize expressed genes through clustering, assembly, and mapping to genomic sequences. However, isolating mRNA from some tissues can be difficult and ESTs alone do not indicate the genes they were derived from.
CRISPR-Cas is a natural defense system in bacteria that uses CRISPR sequences and Cas proteins to target and degrade foreign DNA such as from viruses. It has been adapted for genome editing in other organisms using a Cas9 protein guided by a synthetic single guide RNA to introduce targeted double-strand breaks. This system allows for precise genome modifications and has applications in biomedical research, disease treatment, and engineering of plants and other organisms. However, off-target effects and delivery methods require further optimization.
i explained about basics of genome engineering and crispr system.
CRISPR will change the world and it is just the beginning, are you ready to meet the future? you think its great and beautiful or.....?
please give your feedback to my email
pooyanaghshbandi@yahoo.com
i am starting to write a critical and fantastic review article about CRISPR, if you are interested to join please contact me.
CRISPR-Cas9 is a gene editing technology that uses the bacterial immune system to cut DNA at specific locations. It allows researchers to understand, characterize, and control DNA. CRISPR-Cas9 uses an RNA-guided DNA endonuclease enzyme called Cas9 that is directed by guide RNA to cleave target DNA. It has numerous applications including modifying genes in plants and animals, developing disease resistant crops, and potentially curing genetic diseases in humans by precisely editing genes. While revolutionary, it also raises ethical concerns that must be considered and addressed.
Antisense RNA technology uses single-stranded RNA complementary to messenger RNA to inhibit translation by binding to the mRNA and activating RNase H degradation of the mRNA. While RNAi uses Dicer enzymes, antisense RNA relies on RNase H. The first FDA approved genetically modified food, the Flavr-Savr tomato, used antisense RNA to inhibit the polygalacturonase enzyme and extend tomato shelf life. NIPGR developed tomatoes that could last 45 days using antisense RNA to silence genes responsible for loss of firmness during ripening. Antisense therapy is also being researched to treat diseases by introducing antisense RNA to pathogenic genes.
The document provides an overview of the CRISPR/Cas9 gene editing technology. It discusses the history and components of the CRISPR system, how it works, applications in various fields like microbiology, biomedicine, agriculture, and therapeutics. Recent advances expand its use for transcriptional regulation, epigenetic editing, and live imaging. While powerful, it faces challenges like off-target effects that require further research to optimize its safe and ethical application.
RNAi is a powerful, conserved biological process through which the small, double-stranded RNAs specifically silence the expression of homologous genes, largely through degradation of their cognate mRNA.
Whole genome sequencing is a technique to sequence the entire genome of an organism. It involves breaking the genome into small fragments, copying the fragments, sequencing the fragments, and reassembling the sequence data into the full genome. Key steps include isolating DNA, fragmenting it, ligating fragments into plasmids, amplifying the plasmids, sequencing the fragments using Sanger sequencing, and assembling the sequence reads into the complete genome. Whole genome sequencing allows researchers to discover coding and non-coding regions, predict disease susceptibility, and perform evolutionary studies by comparing species.
RNA interference (RNAi): Cellular process by which an mRNA is targeted for degradation by a dsRNA with a strand complementary to a fragment of such mRNA.
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.
Single strand conformation polymorphismNivethitha T
Single-strand conformation polymorphism (SSCP) is a technique that detects variations in single-stranded DNA sequences. It involves PCR amplification of a target region, denaturing the PCR products to generate single strands, and separating the single strands on a non-denaturing gel based on differences in electrophoretic mobility caused by variations in nucleotide sequence. This allows sequences to be distinguished and variants detected without sequencing. SSCP is useful for discovering new polymorphisms and detecting mutations for diagnostic applications.
The document describes the steps of Illumina sequencing. Genomic DNA is first fragmented and adapters are ligated to create single-stranded DNA fragments. These fragments are attached to a flow cell and undergo bridge amplification to create clusters of identical DNA fragments. Sequencing occurs through cycles of reversible terminator-based sequencing using fluorescently labeled dNTPs, imaging of the fluorescence, and cleavage of the label and terminator to allow the next cycle. After multiple cycles, the sequenced reads are aligned to the reference genome to determine the original sequence.
This presentation discusses strategies for developing transgenic plants without selectable marker genes. Marker genes are commonly used to identify transformed cells but can be problematic for public acceptance and future transformations. Methods described for producing marker-free transgenics include the MAT system which uses oncogenes for selection instead of antibiotics, site-specific recombination systems which flank the marker gene for later excision, and transposon-based systems which separate the gene of interest from the marker gene. While several viable methods exist, more work is still needed before marker-free crops can be commercialized. Removing marker genes supports multiple-gene stacking and improves public acceptance of transgenic technologies.
Dr. Shamalamma S. presented on DNA microarrays. DNA microarrays allow thousands of genes to be compared simultaneously by attaching DNA probes to a chip which fluorescently labeled samples can bind to. The chip is then scanned to analyze gene expression levels. Applications include disease diagnosis, toxicology studies, and pharmacogenomics. While a powerful tool, microarrays have limitations such as lack of knowledge about many genes and lack of standardization.
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.
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.
This document discusses genome editing techniques. It begins by defining genomes and how they consist of DNA or RNA that contains both coding and non-coding regions. It then discusses several methods of genome editing including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR-Cas system. Each method uses engineered nucleases to introduce targeted double-strand breaks in DNA, allowing the cell's repair mechanisms to modify the genome. The CRISPR-Cas system was selected as the breakthrough of the year in 2015 due to its simplicity, efficiency and precision for genome editing applications.
Comparative genomics involves comparing genomes to discover similarities and differences. It can provide insights into evolutionary relationships, help predict gene function, and aid in drug discovery. The first step is often aligning genome sequences using tools like BLAST or MUMmer. Genomes can then be compared at various levels, such as overall nucleotide statistics, genome structure, and coding/non-coding regions. Comparing gene and protein content across genomes helps predict functions. Conserved genomic features across species also aid prediction. Insights into genome evolution come from studying molecular events like inversions and duplications. Comparative genomics has impacted phylogenetics and drug target identification.
Genome editing techniques allow DNA to be inserted, deleted, modified or replaced in the genome of a living organism. Four families of engineered nucleases have been used for genome editing: meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR/Cas9 system. Each system uses a different mechanism for recognizing and cutting DNA at specific locations to modify genes. The CRISPR/Cas9 system has become widely used due to its ease of design and lower off-target effects compared to other techniques.
This document discusses forward and reverse genetic approaches for understanding gene function. Forward genetics begins with a phenotype and identifies the underlying gene, while reverse genetics starts with a gene and determines its phenotype. Specific reverse genetic techniques described include large-scale random mutagenesis, homologous recombination, transposable element excision, RNA interference, genome editing using ZFNs/TALENs/CRISPR, and site-directed mutagenesis combined with transgenics. The document provides details on how each technique is used to alter genes and study their function.
Zinc finger nucleases (ZFNs) allow for highly targeted editing of the genome. ZFNs consist of a DNA-binding domain made of zinc finger proteins and a DNA-cleaving domain. The ZFN pair binds to a target site and creates a double-strand break, which the cell repairs through non-homologous end joining or homologous recombination, enabling gene knockouts or targeted changes. ZFNs work in many cell types and animal models, providing a more efficient alternative to traditional transgenic techniques. They have applications in functional genomics, cell line engineering, and animal model generation.
ESTs are short sequences of DNA derived from cDNA clones that represent gene expression in particular cells or tissues. They provide a simple and inexpensive way to discover new genes and map their positions in genomes. To create an EST, mRNA is converted to cDNA and then sequenced, yielding short expressed DNA sequences. ESTs are deposited in public databases like NCBI's dbEST and can help identify genes, construct genome maps, and characterize expressed genes through clustering, assembly, and mapping to genomic sequences. However, isolating mRNA from some tissues can be difficult and ESTs alone do not indicate the genes they were derived from.
CRISPR-Cas is a natural defense system in bacteria that uses CRISPR sequences and Cas proteins to target and degrade foreign DNA such as from viruses. It has been adapted for genome editing in other organisms using a Cas9 protein guided by a synthetic single guide RNA to introduce targeted double-strand breaks. This system allows for precise genome modifications and has applications in biomedical research, disease treatment, and engineering of plants and other organisms. However, off-target effects and delivery methods require further optimization.
i explained about basics of genome engineering and crispr system.
CRISPR will change the world and it is just the beginning, are you ready to meet the future? you think its great and beautiful or.....?
please give your feedback to my email
pooyanaghshbandi@yahoo.com
i am starting to write a critical and fantastic review article about CRISPR, if you are interested to join please contact me.
CRISPR-Cas9 is a gene editing technology that uses the bacterial immune system to cut DNA at specific locations. It allows researchers to understand, characterize, and control DNA. CRISPR-Cas9 uses an RNA-guided DNA endonuclease enzyme called Cas9 that is directed by guide RNA to cleave target DNA. It has numerous applications including modifying genes in plants and animals, developing disease resistant crops, and potentially curing genetic diseases in humans by precisely editing genes. While revolutionary, it also raises ethical concerns that must be considered and addressed.
Antisense RNA technology uses single-stranded RNA complementary to messenger RNA to inhibit translation by binding to the mRNA and activating RNase H degradation of the mRNA. While RNAi uses Dicer enzymes, antisense RNA relies on RNase H. The first FDA approved genetically modified food, the Flavr-Savr tomato, used antisense RNA to inhibit the polygalacturonase enzyme and extend tomato shelf life. NIPGR developed tomatoes that could last 45 days using antisense RNA to silence genes responsible for loss of firmness during ripening. Antisense therapy is also being researched to treat diseases by introducing antisense RNA to pathogenic genes.
The document provides an overview of the CRISPR/Cas9 gene editing technology. It discusses the history and components of the CRISPR system, how it works, applications in various fields like microbiology, biomedicine, agriculture, and therapeutics. Recent advances expand its use for transcriptional regulation, epigenetic editing, and live imaging. While powerful, it faces challenges like off-target effects that require further research to optimize its safe and ethical application.
RNAi is a powerful, conserved biological process through which the small, double-stranded RNAs specifically silence the expression of homologous genes, largely through degradation of their cognate mRNA.
Whole genome sequencing is a technique to sequence the entire genome of an organism. It involves breaking the genome into small fragments, copying the fragments, sequencing the fragments, and reassembling the sequence data into the full genome. Key steps include isolating DNA, fragmenting it, ligating fragments into plasmids, amplifying the plasmids, sequencing the fragments using Sanger sequencing, and assembling the sequence reads into the complete genome. Whole genome sequencing allows researchers to discover coding and non-coding regions, predict disease susceptibility, and perform evolutionary studies by comparing species.
RNA interference (RNAi): Cellular process by which an mRNA is targeted for degradation by a dsRNA with a strand complementary to a fragment of such mRNA.
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.
Single strand conformation polymorphismNivethitha T
Single-strand conformation polymorphism (SSCP) is a technique that detects variations in single-stranded DNA sequences. It involves PCR amplification of a target region, denaturing the PCR products to generate single strands, and separating the single strands on a non-denaturing gel based on differences in electrophoretic mobility caused by variations in nucleotide sequence. This allows sequences to be distinguished and variants detected without sequencing. SSCP is useful for discovering new polymorphisms and detecting mutations for diagnostic applications.
The document describes the steps of Illumina sequencing. Genomic DNA is first fragmented and adapters are ligated to create single-stranded DNA fragments. These fragments are attached to a flow cell and undergo bridge amplification to create clusters of identical DNA fragments. Sequencing occurs through cycles of reversible terminator-based sequencing using fluorescently labeled dNTPs, imaging of the fluorescence, and cleavage of the label and terminator to allow the next cycle. After multiple cycles, the sequenced reads are aligned to the reference genome to determine the original sequence.
This presentation discusses strategies for developing transgenic plants without selectable marker genes. Marker genes are commonly used to identify transformed cells but can be problematic for public acceptance and future transformations. Methods described for producing marker-free transgenics include the MAT system which uses oncogenes for selection instead of antibiotics, site-specific recombination systems which flank the marker gene for later excision, and transposon-based systems which separate the gene of interest from the marker gene. While several viable methods exist, more work is still needed before marker-free crops can be commercialized. Removing marker genes supports multiple-gene stacking and improves public acceptance of transgenic technologies.
Dr. Shamalamma S. presented on DNA microarrays. DNA microarrays allow thousands of genes to be compared simultaneously by attaching DNA probes to a chip which fluorescently labeled samples can bind to. The chip is then scanned to analyze gene expression levels. Applications include disease diagnosis, toxicology studies, and pharmacogenomics. While a powerful tool, microarrays have limitations such as lack of knowledge about many genes and lack of standardization.
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.
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.
This document provides information on CRISPR Cas9 genome editing. It discusses the history and discovery of CRISPR dating back to 1987. It describes the key components of the CRISPR Cas9 system including Cas9 proteins, CRISPR RNA, protospacers, and PAM sequences. The mechanisms of how CRISPR Cas9 edits genomes through double strand breaks is explained. Finally, applications of CRISPR Cas9 are summarized, including using it to correct genetic mutations causing diseases in animals and potential applications in humans.
Multi Target Gene Editing using CRISPR Technology for Crop ImprovementTushar Gajare
This document provides an overview of a presentation on using CRISPR technology for multi-target gene editing in crop improvement. It begins with an introduction to genome editing and CRISPR-Cas9. It then discusses the CRISPR system, how CRISPR-Cas9 works, its history and applications for crop improvement including case studies in maize with high mutant efficiencies and targeted mutagenesis of multiple genes. The presentation covers advantages and limitations of the technology as well as future prospects.
Crispr cas:an advance and efficient tool for genome modificationavinash tiwari
This document provides an overview of CRISPR-Cas as an advanced and efficient tool for genome modification. It describes how CRISPR-Cas systems incorporate DNA from invading viruses or plasmids and use RNA-guided Cas nucleases to cleave matching sequences in foreign DNA. The two main components required for genome editing are Cas9 nuclease and a guide RNA. By altering the guide RNA sequence, Cas9 can be directed to cleave any desired DNA target. The document discusses applications of CRISPR-Cas in genome editing, gene regulation, molecular barcoding, and potential future uses in medicine and biotechnology.
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
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.
Genome editing with engineered nucleasesKrishan Kumar
Genome editing uses engineered nucleases to insert, replace or remove DNA from the genome. These nucleases create targeted double-strand breaks which are repaired through natural DNA repair processes, allowing for changes to the genome sequence. Three main engineered nuclease systems for genome editing are ZFNs, TALENs, and CRISPR-Cas9. CRISPR uses a guide RNA and Cas9 nuclease to make precise cuts at targeted DNA sequences for editing. It has advantages over ZFNs and TALENs in being cheaper, easier to design, and more efficient. Genome editing holds promise for applications in crops, medicine, and research.
Recent advances in CRISPR-CAS9 technology: an alternative to transgenic breedingJyoti Prakash Sahoo
These are the part of the Bacterial immune system which detects and recognize the foreign DNA and cleaves it.
THE CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) loci
Cas (CRISPR- associated) proteins can target and cleave invading DNA in a sequence – specific manner.
CRISPR array is composed of a series of repeats interspaced by spacer sequences acquired from invading genomes.
CRISPR/Cas9 is an advanced genome editing technology that can be used to develop plant disease resistance. It involves a Cas9 enzyme that acts like molecular scissors to cut DNA at specific locations guided by CRISPR RNA. This triggers DNA repair that can introduce changes to genes. Researchers have used CRISPR/Cas9 to develop resistance in plants against viruses, fungi, and bacteria by editing genes involved in host-pathogen interaction and disease susceptibility. It provides a precise and efficient way to edit plant genomes to improve crop resistance compared to previous tools. Scientists continue working to enhance the specificity and control of CRISPR/Cas9 for genome editing applications in agriculture.
This document provides an overview of CRISPR/Cas9 genome editing. It discusses how CRISPR/Cas9 enables precise modification of DNA sequences, outlines the timeline of key discoveries in CRISPR research, and describes the molecular mechanism and potential applications of this technology, including in microbial research, crop improvement, and human gene therapy. It also notes some limitations of the CRISPR/Cas9 system and concludes by emphasizing the opportunities it provides to advance research and address challenges like food security.
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.
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
This document summarizes a presentation on CRISPR/Cas genome editing. It defines CRISPR/Cas as a type of genetic engineering that uses artificially engineered nucleases to make specific cuts in DNA. It describes the CRISPR/Cas system's origins and components, including Cas9, guide RNA, and PAM sequences. Applications discussed include genome editing in animals and plants, as well as concerns over off-target effects. Companies offering CRISPR services or kits are also mentioned.
CRISPR-Cas is an adaptive immune system existing in most bacteria and archaea, preventing them from being infected by phages, viruses and other foreign genetic elements.
This presentation explains about the working and applications of CRISPR-CAS system.
Genome engineering using CRISPR/Cas9 has several advantages over traditional gene targeting methods: it is faster, more precise, applicable to many species, and less expensive. CRISPR/Cas9 uses the Cas9 nuclease guided by a single guide RNA to introduce double-strand breaks at targeted genomic loci. This can generate gene knockouts through error-prone non-homologous end joining or allow for targeted insertions and modifications through homology-directed repair. While CRISPR/Cas9 has great potential, careful design of guide RNAs and donor templates is needed to minimize off-target effects.
A review of the growth of the Israel Genealogy Research Association Database Collection for the last 12 months. Our collection is now passed the 3 million mark and still growing. See which archives have contributed the most. See the different types of records we have, and which years have had records added. You can also see what we have for the future.
Assessment and Planning in Educational technology.pptxKavitha Krishnan
In an education system, it is understood that assessment is only for the students, but on the other hand, the Assessment of teachers is also an important aspect of the education system that ensures teachers are providing high-quality instruction to students. The assessment process can be used to provide feedback and support for professional development, to inform decisions about teacher retention or promotion, or to evaluate teacher effectiveness for accountability purposes.
This presentation includes basic of PCOS their pathology and treatment and also Ayurveda correlation of PCOS and Ayurvedic line of treatment mentioned in classics.
This presentation was provided by Steph Pollock of The American Psychological Association’s Journals Program, and Damita Snow, of The American Society of Civil Engineers (ASCE), for the initial session of NISO's 2024 Training Series "DEIA in the Scholarly Landscape." Session One: 'Setting Expectations: a DEIA Primer,' was held June 6, 2024.
Strategies for Effective Upskilling is a presentation by Chinwendu Peace in a Your Skill Boost Masterclass organisation by the Excellence Foundation for South Sudan on 08th and 09th June 2024 from 1 PM to 3 PM on each day.
A workshop hosted by the South African Journal of Science aimed at postgraduate students and early career researchers with little or no experience in writing and publishing journal articles.
ISO/IEC 27001, ISO/IEC 42001, and GDPR: Best Practices for Implementation and...PECB
Denis is a dynamic and results-driven Chief Information Officer (CIO) with a distinguished career spanning information systems analysis and technical project management. With a proven track record of spearheading the design and delivery of cutting-edge Information Management solutions, he has consistently elevated business operations, streamlined reporting functions, and maximized process efficiency.
Certified as an ISO/IEC 27001: Information Security Management Systems (ISMS) Lead Implementer, Data Protection Officer, and Cyber Risks Analyst, Denis brings a heightened focus on data security, privacy, and cyber resilience to every endeavor.
His expertise extends across a diverse spectrum of reporting, database, and web development applications, underpinned by an exceptional grasp of data storage and virtualization technologies. His proficiency in application testing, database administration, and data cleansing ensures seamless execution of complex projects.
What sets Denis apart is his comprehensive understanding of Business and Systems Analysis technologies, honed through involvement in all phases of the Software Development Lifecycle (SDLC). From meticulous requirements gathering to precise analysis, innovative design, rigorous development, thorough testing, and successful implementation, he has consistently delivered exceptional results.
Throughout his career, he has taken on multifaceted roles, from leading technical project management teams to owning solutions that drive operational excellence. His conscientious and proactive approach is unwavering, whether he is working independently or collaboratively within a team. His ability to connect with colleagues on a personal level underscores his commitment to fostering a harmonious and productive workplace environment.
Date: May 29, 2024
Tags: Information Security, ISO/IEC 27001, ISO/IEC 42001, Artificial Intelligence, GDPR
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How to Fix the Import Error in the Odoo 17Celine George
An import error occurs when a program fails to import a module or library, disrupting its execution. In languages like Python, this issue arises when the specified module cannot be found or accessed, hindering the program's functionality. Resolving import errors is crucial for maintaining smooth software operation and uninterrupted development processes.
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Physiology and chemistry of skin and pigmentation, hairs, scalp, lips and nail, Cleansing cream, Lotions, Face powders, Face packs, Lipsticks, Bath products, soaps and baby product,
Preparation and standardization of the following : Tonic, Bleaches, Dentifrices and Mouth washes & Tooth Pastes, Cosmetics for Nails.
Exploiting Artificial Intelligence for Empowering Researchers and Faculty, In...Dr. Vinod Kumar Kanvaria
Exploiting Artificial Intelligence for Empowering Researchers and Faculty,
International FDP on Fundamentals of Research in Social Sciences
at Integral University, Lucknow, 06.06.2024
By Dr. Vinod Kumar Kanvaria
How to Build a Module in Odoo 17 Using the Scaffold MethodCeline George
Odoo provides an option for creating a module by using a single line command. By using this command the user can make a whole structure of a module. It is very easy for a beginner to make a module. There is no need to make each file manually. This slide will show how to create a module using the scaffold method.
How to Build a Module in Odoo 17 Using the Scaffold Method
Crisper Cas system
1. CRISPR cas : A new genome editing tool
Presented by
Surbhaiyya shobha devidas
Ph.D Scholar
Dr. PDKV, Akola
1
2. 1. Introduction
2. History
3. CRISPR in bacteria
4. Classification of CRISPR
5. General structure of cas9 protein
6. Mechanism of CRISPR cas9
7. Applications
8. Data base of CRISPR
9. Case study
10. Conclusion
11. Future aspects
Content…
3. • Genome editing, or genome editing with engineered nucleases (GEEN) is a type
of genetic engineering in which DNA is inserted, replaced, or removed from
a genome using artificially engineered nucleases, or "molecular scissors”.
• The nucleases create specific double-strand breaks (DSBs) at desired locations in the
genome and harness the cell’s endogenous mechanisms to repair the induced break by
natural processes of homologous recombination (HR) and non-homologous end-
joining (NHEJ).
3
introduction…
4. To understand the function of a gene or a protein, one interferes with it in a sequence-specific way and
monitors its effects on the organism.
In some organisms, it is difficult or impossible to perform site-specific mutagenesis, and therefore more
indirect methods must be used, such as silencing the gene of interest by short RNA interference (siRNA).
But sometime gene disruption by siRNA can be variable or incomplete.
Nucleases such as CRISPR can cut any targeted position in the genome and introduce a modification of the
endogenous sequences for genes that are impossible to specifically target using conventional RNAi.
4
Why genome editing?
6. Comparison between traditional and modern genome editing
technologies
Mutagen Chemical(e.g., EMS) Physical (e.g.,
gamma, X-ray or fast
neutron radiation)
Biological (ZFNs,
TALENs or CRISPR/
Cas)
Biological-
Transgenics (e.g.,
Agro or gene gun)
Characteristi
cs of genetic
variation
Substitution and Deletion Deletion and
chromosomal
mutation
Substitution and
Deletion and
insertion
Insertions
Loss of function Loss of function Loss of function and
gain of function
Loss of function and
gain of function
Advantages Unnecessary of knowing
gene function or sequences
Unnecessary of
knowing gene
function or
sequences
Gene specific
mutation
Insertion of genes of
known functions
into host plant
genome
Easy production of random
mutation
Easy production of
random mutation
Efficient production
of desirable
mutation
Efficient creation of
plants with desirable
traits 6
7. Disadvantages Inefficient screening of
desirable traits
Inefficient screening
of desirable traits
Necessity of knowing
gene function and
sequences
Necessity of knowing
gene function and
sequences
Non specific mutation Non specific
mutation
Prerequisite of
efficient genetic
transformation
Prerequisite of
efficient genetic
transformation
Other features Non transgenic process
and traits
Non transgenic
process and traits
Transgenic process
but non transgenic
traits
Transgenic process
and traits
7
8. 1987
• Researchers find CRISPR sequences in Escherichia coli, but do not
characterize their function.
2000
• CRISPR sequence are found to be common in other microbes.
2002
• Coined CRISPR name, defined signature Cas genes
2007
• First experimental evidence for CRISPR adaptive immunity
2013
• First demonstration of Cas9 genome engineering in eukaryotic cell
HISTORY
8
9. CRISPR – Cas systems
• These are the part of the Bacterial immune system which detects
and recognize the foreign DNA and cleaves it.
1. THE CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)
loci
2. Cas (CRISPR- associated) proteins can target and cleave invading DNA in a
sequence – specific manner.
A CRISPR array is composed of a series of repeats interspaced by
spacer sequences acquired from invading genomes.
9
11. Different CRISPR-Cas system in Bacterial Adaptive Immunity
Class 1- type I (CRISPR-Cas3) and type III (CRISPR-
Cas10)
uses several Cas proteins and the crRNA
Class 2- type II (CRISPR-Cas9) and type V (CRISPR-
Cpf1)
employ a large single-component Cas-9 protein in
conjunction with crRNA and tracerRNA.
11Zetsche et al., (2015)
functioning of type II CRISPR system
12. Different Cas proteins and their function
Protein Distribution Process Function
Cas1 Universal Spacer acquisition DNAse, not sequence specfic, can bind RNA; present in all Types
Cas2 Universal Spacer acquisition specific to U-rich regions; present in all Types
Cas3 Type I signature Target interference DNA helicase, endonuclease
Cas4 Type I, II Spacer acquisition RecB-like nuclease with exonuclease activity homologous to RecB
Cas5 Type I crRNA expression RAMP protein, endoribonuclease involved in crRNA biogenesis; part of CASCADE
Cas6 Type I, III crRNA expression RAMP protein, endoribonuclease involved in crRNA biogenesis; part of CASCADE
Cas7 Type I crRNA expression RAMP protein, endoribonuclease involved in crRNA biogenesis; part of CASCADE
Cas8 Type I crRNA expression Large protein with McrA/HNH-nuclease domain and RuvC-like nuclease; part of
CASCADE
Cas9 Type II signature Target interference Large multidomain protein with McrA-HNH nuclease domain and RuvC-like
nuclease domain; necessary for interference and target cleavage
Cas10 Type III signature crRNA expression
and interference
HD nuclease domain, palm domain, Zn ribbon; some homologies with CASCADE
elements
12
13. Action of CRISPR in bacteria
The CRISPR immune system works to protect bacteria from
repeated viral attack via three basic steps:
(1) Adaptation
(2) Production of crRNA
(3) Targeting
13
29. Cpf1 (CRISPR from Prevotella and Francisella 1) at Broad Institute of MIT and
Harvard, Cambridge.
CRISPR-Cpf1 is a class 2 CRISPR system
Does not require tracerRNA and the gene is 1kb smaller
Targeted DNA is cleaved as a 5 nt staggered cut distal to a 5’ T-rich PAM
Cpf1 exhibit robust nuclease activity in human cells
29
Zetsche et al., (October 22, 2015)
New Version of Cas9:
30. 30
Organization of two CRISPR loci found in Francisella novicida .The domain architectures
of FnCas9 and FnCpf1 are compared
31. Application in Agriculture
Can be used to create high degree of genetic variability at precise locus in the
genome of the crop plants.
Potential tool for multiplexed reverse and forward genetic study.
Precise transgene integration at specific loci.
Developing biotic and abiotic resistant traits in crop plants.
Potential tool for developing virus resistant crop varieties.
Can be used to eradicate unwanted species like herbicide resistant weeds,
insect pest.
Potential tool for improving polyploidy crops like potato and wheat.
31
32. sgRNA designing tools
Optimized CRISPR Design (Feng Zhang's Lab at MIT/BROAD, USA)
sgRNA Scorer (George Church's Lab at Harvard, USA)
sgRNA Designer (BROAD Institute)
ChopChop web tool (George Church's Lab at Harvard, USA)
E-CRISP (Michael Boutros' lab at DKFZ, Germany)
CRISPR Finder (Wellcome Trust Sanger Institute, Hinxton, UK)
RepeatMasker (Institute for Systems Biology) to double check and avoid selecting target sites
with repeated sequences
32
37. MATERIALS AND METHODS
• Plant material – Soybean line
• Plasmid construction of cas9 and sgRNA
Two GFP-targeting gRNA vectors were designed;
one gRNA was designed to target the 5′ end of GFP (5′- target) and a second was
designed to target the 3′ end (3′-target)
• Hairy root transformation of soybean by A. rhizogenes (strain K599)
• GFP imaging via Olympus MVX10 microscope with a GFP filter
• sequencing and analysis
37
39. (B)C9 + GFP 5' target events
• Wild-type sequences are in green, deletions are shown as
dashes, and SNPs are shown in orange.
40. (C) C9 + GFP 3' target events
• Wild-type sequences are in green, deletions are shown as dashes,
and SNPs are shown in orange.
41. Conclusion…
Genome editing tools provide new strategies for genetic manipulation in
plants and are likely to assist in engineering desired plant traits by
modifying endogenous genes.
Genome editing technology will have a major impact in applied crop
improvement and commercial product development .
In gene modification, these targetable nucleases have potential
applications to become alternatives to standard breeding methods to
identify novel traits in economically important plants and more valuable in
biotechnology as modifying specific site rather than whole gene.
41