RNA interference & its application
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RNA interference is a mechanism that inhibits gene expression at the translation stage or hinders gene transcription. It is described by three similar but phenotypically different phenomena: co-suppression in plants, quelling in fungi, and RNA interference in animals. Recent applications of RNA interference include modulating HIV replication, inhibiting HIV, treating hepatitis, intervening in atherosclerosis, and reducing heart and brain damage. RNA interference has diverse therapeutic applications including identifying gene functions, creating genome-wide screens, developing antiviral and anticancer therapies.
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RNA Interference technology
RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression, typically by causing the destruction of specific mRNA molecules. Historically, it was known by other names, including co-suppression, post-transcriptional gene silencing (PTGS), and quelling.
Oligonucleotide ligation assay
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.
SiRNA
Kashikant Yadav presented on siRNA (short interfering RNA). siRNA is 20-25 base pairs long, similar to miRNA, and operates in the RNA interference pathway by degrading mRNA with complementary sequences, preventing translation. There are three main methods of siRNA synthesis: chemical synthesis, in vitro transcription, and digestion of long dsRNA by RNAase III or Dicer. siRNA has significance for protecting against viruses, maintaining genome stability, and offers a new tool to specifically repress genes. Potential applications include testing gene function, target validation, pathway analysis, and developing siRNA therapeutics.
Genomics and Plant Genomics
Genomics refers to the study of the entire genome of an organism. It deals with mapping genes on chromosomes and sequencing entire genomes. While work on genomics began with prokaryotes like bacteria, research has now been conducted on crop plants like rice and Arabidopsis thaliana. Genomics is an interdisciplinary field that uses tools from molecular biology, robotics, and computing to study genomes. It provides information on genome size, gene number, gene function, and evolution. Genomics has applications in crop improvement through gene mapping, marker-assisted selection, and transgenic breeding. However, genomic research also faces limitations due to high costs, technical challenges, and complexity of traits.
Gene silencing and its significance
Gene silencing is a molecular process that down regulates gene expression at the transcriptional or translational level. It allows researchers to study essential genes and provides insight into disease development since many diseases are associated with reduced gene expression. Gene silencing occurs through various mechanisms including histone modification, DNA methylation, and small RNA pathways. It has applications in research areas like producing virus resistant plants and modifying food quality and plant traits.
Rna interference
RNA interference (RNAi) is a biological mechanism that leads to post-transcriptional gene silencing triggered by double-stranded RNA (dsRNA) molecules. It involves small interfering RNAs (siRNAs) and microRNAs (miRNAs) that are processed by Dicer nuclease and loaded into an RNA-induced silencing complex (RISC) which targets and degrades messenger RNAs (mRNAs). The mechanism of RNAi involves dsRNA being cleaved by Dicer into siRNAs which are incorporated into RISC and used as a guide to degrade complementary mRNAs. RNAi has various applications in crop improvement traits like enhanced shelf life, male sterility/fertility, biofortification, aller
RNA interference
This document discusses RNA interference (RNAi) and its mechanisms. It can be summarized as follows:
1. RNAi is a process where double-stranded RNA causes degradation of homologous mRNA sequences. It was discovered in 1998 and is found across many organisms.
2. The RNAi pathway involves conversion of dsRNA to siRNAs by the enzyme Dicer. siRNAs are incorporated into the RISC complex containing Argonaute proteins. RISC then cleaves and destroys homologous mRNA targets.
3. miRNAs are endogenous single-stranded RNAs that regulate gene expression at the translation level by preventing ribosome binding. They are processed from hairpin precursors by the enzymes Dro
RNA interference (RNAi)
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.
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RNA Interference technology
RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression, typically by causing the destruction of specific mRNA molecules. Historically, it was known by other names, including co-suppression, post-transcriptional gene silencing (PTGS), and quelling.
Oligonucleotide ligation assay
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.
SiRNA
Kashikant Yadav presented on siRNA (short interfering RNA). siRNA is 20-25 base pairs long, similar to miRNA, and operates in the RNA interference pathway by degrading mRNA with complementary sequences, preventing translation. There are three main methods of siRNA synthesis: chemical synthesis, in vitro transcription, and digestion of long dsRNA by RNAase III or Dicer. siRNA has significance for protecting against viruses, maintaining genome stability, and offers a new tool to specifically repress genes. Potential applications include testing gene function, target validation, pathway analysis, and developing siRNA therapeutics.
Genomics and Plant Genomics
Genomics refers to the study of the entire genome of an organism. It deals with mapping genes on chromosomes and sequencing entire genomes. While work on genomics began with prokaryotes like bacteria, research has now been conducted on crop plants like rice and Arabidopsis thaliana. Genomics is an interdisciplinary field that uses tools from molecular biology, robotics, and computing to study genomes. It provides information on genome size, gene number, gene function, and evolution. Genomics has applications in crop improvement through gene mapping, marker-assisted selection, and transgenic breeding. However, genomic research also faces limitations due to high costs, technical challenges, and complexity of traits.
Gene silencing and its significance
Gene silencing is a molecular process that down regulates gene expression at the transcriptional or translational level. It allows researchers to study essential genes and provides insight into disease development since many diseases are associated with reduced gene expression. Gene silencing occurs through various mechanisms including histone modification, DNA methylation, and small RNA pathways. It has applications in research areas like producing virus resistant plants and modifying food quality and plant traits.
Rna interference
RNA interference (RNAi) is a biological mechanism that leads to post-transcriptional gene silencing triggered by double-stranded RNA (dsRNA) molecules. It involves small interfering RNAs (siRNAs) and microRNAs (miRNAs) that are processed by Dicer nuclease and loaded into an RNA-induced silencing complex (RISC) which targets and degrades messenger RNAs (mRNAs). The mechanism of RNAi involves dsRNA being cleaved by Dicer into siRNAs which are incorporated into RISC and used as a guide to degrade complementary mRNAs. RNAi has various applications in crop improvement traits like enhanced shelf life, male sterility/fertility, biofortification, aller
RNA interference
This document discusses RNA interference (RNAi) and its mechanisms. It can be summarized as follows:
1. RNAi is a process where double-stranded RNA causes degradation of homologous mRNA sequences. It was discovered in 1998 and is found across many organisms.
2. The RNAi pathway involves conversion of dsRNA to siRNAs by the enzyme Dicer. siRNAs are incorporated into the RISC complex containing Argonaute proteins. RISC then cleaves and destroys homologous mRNA targets.
3. miRNAs are endogenous single-stranded RNAs that regulate gene expression at the translation level by preventing ribosome binding. They are processed from hairpin precursors by the enzymes Dro
RNA interference (RNAi)
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.
RNA interference in Biology and medicine
RNA interference (RNAi) is a biological process where RNA molecules inhibit gene expression, typically by causing the destruction of specific mRNA molecules. It was first discovered in plants and nematodes but is found in most eukaryotic organisms. The process involves specialized enzymes that cut long RNA molecules into short interfering RNAs which then guide other proteins to destroy any RNAs of the same sequence. RNAi plays important roles in regulating genes and defending against foreign DNA and RNA. It has many applications for studying gene function and as a potential therapeutic approach for diseases like cancer and infections.
SNP Detection Methods and applications
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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
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Sequence Polymorphism-Derived (SPD) markers
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Minisequencing primers
Allele-specific ligation probes
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It has various function in Gene regulation and silencing.
SAGE (Serial analysis of Gene Expression)
SAGE (Serial Analysis of Gene Expression) is a technique that allows for the rapid and comprehensive analysis of gene expression patterns in a given cell population. It works by isolating mRNA, synthesizing cDNA, ligating short sequence tags to the cDNA, and then counting the number of times each tag is observed to quantify gene expression levels. The tags are concatenated and sequenced to generate vast amounts of data that must be analyzed computationally to identify which genes particular tags correspond to and to compare expression profiles between cell types. SAGE provides an overview of a cell's complete transcriptional activity and has been applied to study differences in cancer vs normal cells and to identify targets of oncogenes and tumor suppressor genes.
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Maxam gilbert sequencing method
The Maxam-Gilbert sequencing method involves chemically cleaving DNA strands at specific bases, labeling the fragments radioactively, and separating them via gel electrophoresis. The sequence can then be read from the banding pattern on X-ray film. This early chemical degradation method was developed in 1977 and provided one of the first reliable ways to determine DNA sequences, but it uses toxic chemicals and radioactivity, has technical complexity, and can only read up to 500 base pairs.
Gene sequencing methods
This document discusses various gene sequencing methods. It begins by introducing DNA and the importance of sequencing the genetic code. It then describes several early sequencing techniques like Sanger sequencing using chain termination or chemical cleavage. It discusses the need for sequencing to understand genetic conditions. The document also covers topics like genome sequencing, genomics, and high-throughput sequencing techniques like dye-terminator sequencing which replaced radioactive labels with fluorescent labels to automate the process.
Micro RNAs
This document discusses microRNAs (miRNAs), which are 22 nucleotide non-coding RNAs that play important regulatory roles in plants. miRNAs are processed from stem-loop precursors by the enzyme Dicer and mediate post-transcriptional gene silencing by guiding mRNA cleavage or translational repression. Bioinformatic and genetic studies have identified many conserved miRNA families in plants and shown that miRNAs regulate key transcription factors to control developmental processes.
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anti sense RNA technology is one the method for gene silencing. it found very effective making Flavr savr tomato. hope this ppt will help you.
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This document discusses RNA interference (RNAi), a biological mechanism that leads to post-transcriptional gene silencing triggered by double-stranded RNA molecules. It was discovered in 1998 by Fire and Mello, who received the Nobel Prize for this work. The mechanism involves double-stranded RNA being processed by the enzyme Dicer into small interfering RNAs that integrate into the RNA-induced silencing complex and guide mRNA degradation. RNAi can be induced by synthetic siRNAs or endogenous microRNAs and represents a powerful tool for studying gene function and developing therapies.
Sanger sequencing (DNA sequencing by ENZYMATIC METHOD)
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Aflp (amplified fragment length polymorphism), alu
Alu PCR amplifies repetitive Alu elements in DNA using Alu-specific primers. The presence or absence of Alu insertions can be used for population genetics, forensics, and paternity testing. Alu elements are short, inter
Zinc Finger Nuclease.
This document provides information about zinc finger proteins. It begins with an introduction to zinc finger motifs, which are protein structural domains characterized by the coordination of zinc ions. The document then discusses the history of zinc finger discovery, functions, and families. It provides details on the most common Cys2His2 zinc finger proteins and their role in DNA recognition and transcriptional regulation. The document also examines uses of zinc finger nucleases for genome editing and their mechanism of action involving creating double-strand breaks in DNA.
Molecular tagging
Molecular tagging of genes involves identifying existing DNA or introducing new DNA to function as a tag or label for the gene of interest. There are four main strategies for gene tagging: marker-based tagging, transposon tagging, T-DNA tagging, and epitope tagging. Marker-based tagging uses molecular markers tightly linked to important traits to assist in plant breeding programs. Transposon tagging relies on transposons, which can move within the genome, to provide a DNA tag that can then be used to identify adjacent DNA sequences and genes.
Dna sequencing
1) DNA sequencing refers to determining the order of nucleotide bases (A, G, C, T) in a DNA molecule. This provides essential genetic information for growth and development.
2) Two major early methods for DNA sequencing were the chemical cleavage method developed by Maxam and Gilbert in 1977 and the chain termination method developed by Sanger. Sanger's method became more popular due to fewer toxic chemicals.
3) Modern DNA sequencing often uses fluorescent dye-labeled chain terminators and capillary electrophoresis. Each dye fluoresces at a different wavelength, allowing all four reactions to occur in one tube. This high-throughput automated approach has accelerated genomic research.
cisgenesis and intragenesis
it cover almost all content in cis/intragesis, right from introduction definition, explanation, production of marker free transgenic, intragenic vector construction, regulatory guide lines, current and future status, limitation, advantage over existing technique, swot analysis etc
its very useful for your seminar and presentations. it contain lot of picture, table, figure for your easy understanding
thank you
Mahesh
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This document provides an overview of functional genomics and methods for transcriptome analysis. It discusses two main approaches - sequence-based approaches like expressed sequence tags (ESTs) and serial analysis of gene expression (SAGE), and microarray-based approaches. For sequence-based approaches, it describes how ESTs can provide gene discovery and expression information but have limitations. It outlines the SAGE methodology and gene index construction to organize EST data. For microarrays, it summarizes the basic workflow including sample preparation, hybridization, image analysis and data normalization to identify differentially expressed genes through statistical tests.
TILLING & ECO-TILLING
A new era of genomics for plant science research has opened due the complete genome sequencing projects of Arabidopsis thaliana and rice. The sequence information available in public database has highlighted the need to develop genome scale reverse genetic strategies for functional analysis (Till et al., 2003). As most of the phenotypes are obscure, the forward genetics can hardly meet the demand of a high throughput and large-scale survey of gene functions. Targeting Induced Local Lesions in Genome TILLING is a general reverse genetic technique that combines chemical mutagenesis with PCR based screening to identity point mutations in regions of interest (McCallum et al., 2000). This strategy works with a mismatch-specific endonuclease to detect induced or natural DNA polymorphisms in genes of interest. A newly developed general reverse genetic strategy helps to locate an allelic series of induced point mutations in genes of interest. It allows the rapid and inexpensive detection of induced point mutations in populations of physically or chemically mutagenized individuals. To create an induced population with the use of physical/chemical mutagens is the first prerequisite for TILLING approach. Most of the plant species are compatible with this technique due to their self-fertilized nature and the seeds produced by these plants can be stored for long periods of time (Borevitz et al., 2003). The seeds are treated with mutagens and raised to harvest M1 plants, which are consequently, self-fertilized to raise the M2 population. DNA extracted from M2 plants is used in mutational screening (Colbert et al., 2001). To avoid mixing of the same mutation only one M2 plant from each M1 is used for DNA extraction (Till et al., 2007). The M3 seeds produce by selfing the M2 progeny can be well preserved for long term storage. Ethyl methane sulfonate (EMS) has been extensively used as a chemical mutagen in TILLING studies in plants to generate mutant populations, although other mutagens can be effective. EMS produces transitional mutations (G/C, A/T) by alkylating G residues which pairs with T instead of the conservative base pairing with C (Nagy et al., 2003). It is a constructive approach for users to attempt a range of chemical mutagens to assess the lethality and sterility on germinal tissue before creating large mutant populations.
Presentation1 Gene silencing.pptx
This seminar presentation discusses gene silencing. It introduces gene silencing as the regulation of gene expression to prevent expression of a certain gene, which can occur during transcription or translation. It then describes the main types of gene silencing - transcriptional, post-transcriptional, and meiotic gene silencing. Research methods like antisense oligonucleotides and RNA interference are explained. Applications of gene silencing in cancer, respiratory diseases, and Huntington's disease are provided. The conclusion covers that gene silencing was first observed in plants/fungi but also exists in mammals, and can be used to extinguish expression of harmful genes.
Gene silensing
Gene silencing is a process where the expression of a gene is reduced or turned off. It occurs through transcriptional or post-transcriptional mechanisms. At the transcriptional level, gene expression is modified directly. At the post-transcriptional level, double stranded RNA suppresses gene expression by binding to mRNA and preventing protein production. There are three main gene silencing mechanisms - antisense oligonucleotides, ribozymes, and RNA interference. RNAi has become the most widely used mechanism due to its effectiveness and specificity. Gene silencing has many applications including developing treatments for diseases like cancer, HIV, and respiratory illnesses. It is also used to modify crop traits and study unknown gene functions.
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RNA interference in Biology and medicine
RNA interference (RNAi) is a biological process where RNA molecules inhibit gene expression, typically by causing the destruction of specific mRNA molecules. It was first discovered in plants and nematodes but is found in most eukaryotic organisms. The process involves specialized enzymes that cut long RNA molecules into short interfering RNAs which then guide other proteins to destroy any RNAs of the same sequence. RNAi plays important roles in regulating genes and defending against foreign DNA and RNA. It has many applications for studying gene function and as a potential therapeutic approach for diseases like cancer and infections.
SNP Detection Methods and applications
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
Short hairpin rna
Short Hair Pin RNA is a molecule which takes part in RNA interference technology.
It has various function in Gene regulation and silencing.
SAGE (Serial analysis of Gene Expression)
SAGE (Serial Analysis of Gene Expression) is a technique that allows for the rapid and comprehensive analysis of gene expression patterns in a given cell population. It works by isolating mRNA, synthesizing cDNA, ligating short sequence tags to the cDNA, and then counting the number of times each tag is observed to quantify gene expression levels. The tags are concatenated and sequenced to generate vast amounts of data that must be analyzed computationally to identify which genes particular tags correspond to and to compare expression profiles between cell types. SAGE provides an overview of a cell's complete transcriptional activity and has been applied to study differences in cancer vs normal cells and to identify targets of oncogenes and tumor suppressor genes.
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Genomics, proteomics and metabolomics are the three core omics technologies, which respectively deal with the analysis of genome, proteome and metabolome of cells and tissues of an organism.
mi RNA and siRNA
This document provides an overview of siRNA and miRNA. It defines siRNA as short interfering RNA that is 20-25 base pairs long and similar to miRNA. miRNA is defined as a non-coding RNA molecule around 21-23 nucleotides that inhibits mRNA expression. Both siRNA and miRNA operate in the RNA interference pathway by being processed by the enzyme Dicer and interfering with gene expression by degrading complementary mRNA. The document also reviews the mechanisms and significance of RNAi, including its role in protecting against viruses, maintaining genome stability, and offering a new experimental tool to repress genes specifically.
Maxam gilbert sequencing method
The Maxam-Gilbert sequencing method involves chemically cleaving DNA strands at specific bases, labeling the fragments radioactively, and separating them via gel electrophoresis. The sequence can then be read from the banding pattern on X-ray film. This early chemical degradation method was developed in 1977 and provided one of the first reliable ways to determine DNA sequences, but it uses toxic chemicals and radioactivity, has technical complexity, and can only read up to 500 base pairs.
Gene sequencing methods
This document discusses various gene sequencing methods. It begins by introducing DNA and the importance of sequencing the genetic code. It then describes several early sequencing techniques like Sanger sequencing using chain termination or chemical cleavage. It discusses the need for sequencing to understand genetic conditions. The document also covers topics like genome sequencing, genomics, and high-throughput sequencing techniques like dye-terminator sequencing which replaced radioactive labels with fluorescent labels to automate the process.
Micro RNAs
This document discusses microRNAs (miRNAs), which are 22 nucleotide non-coding RNAs that play important regulatory roles in plants. miRNAs are processed from stem-loop precursors by the enzyme Dicer and mediate post-transcriptional gene silencing by guiding mRNA cleavage or translational repression. Bioinformatic and genetic studies have identified many conserved miRNA families in plants and shown that miRNAs regulate key transcription factors to control developmental processes.
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anti sense RNA technology is one the method for gene silencing. it found very effective making Flavr savr tomato. hope this ppt will help you.
RNA interference
This document discusses RNA interference (RNAi), a biological mechanism that leads to post-transcriptional gene silencing triggered by double-stranded RNA molecules. It was discovered in 1998 by Fire and Mello, who received the Nobel Prize for this work. The mechanism involves double-stranded RNA being processed by the enzyme Dicer into small interfering RNAs that integrate into the RNA-induced silencing complex and guide mRNA degradation. RNAi can be induced by synthetic siRNAs or endogenous microRNAs and represents a powerful tool for studying gene function and developing therapies.
Sanger sequencing (DNA sequencing by ENZYMATIC METHOD)
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Aflp (amplified fragment length polymorphism), alu
Alu PCR amplifies repetitive Alu elements in DNA using Alu-specific primers. The presence or absence of Alu insertions can be used for population genetics, forensics, and paternity testing. Alu elements are short, inter
Zinc Finger Nuclease.
This document provides information about zinc finger proteins. It begins with an introduction to zinc finger motifs, which are protein structural domains characterized by the coordination of zinc ions. The document then discusses the history of zinc finger discovery, functions, and families. It provides details on the most common Cys2His2 zinc finger proteins and their role in DNA recognition and transcriptional regulation. The document also examines uses of zinc finger nucleases for genome editing and their mechanism of action involving creating double-strand breaks in DNA.
Molecular tagging
Molecular tagging of genes involves identifying existing DNA or introducing new DNA to function as a tag or label for the gene of interest. There are four main strategies for gene tagging: marker-based tagging, transposon tagging, T-DNA tagging, and epitope tagging. Marker-based tagging uses molecular markers tightly linked to important traits to assist in plant breeding programs. Transposon tagging relies on transposons, which can move within the genome, to provide a DNA tag that can then be used to identify adjacent DNA sequences and genes.
Dna sequencing
1) DNA sequencing refers to determining the order of nucleotide bases (A, G, C, T) in a DNA molecule. This provides essential genetic information for growth and development.
2) Two major early methods for DNA sequencing were the chemical cleavage method developed by Maxam and Gilbert in 1977 and the chain termination method developed by Sanger. Sanger's method became more popular due to fewer toxic chemicals.
3) Modern DNA sequencing often uses fluorescent dye-labeled chain terminators and capillary electrophoresis. Each dye fluoresces at a different wavelength, allowing all four reactions to occur in one tube. This high-throughput automated approach has accelerated genomic research.
cisgenesis and intragenesis
it cover almost all content in cis/intragesis, right from introduction definition, explanation, production of marker free transgenic, intragenic vector construction, regulatory guide lines, current and future status, limitation, advantage over existing technique, swot analysis etc
its very useful for your seminar and presentations. it contain lot of picture, table, figure for your easy understanding
thank you
Mahesh
Functional genomics
This document provides an overview of functional genomics and methods for transcriptome analysis. It discusses two main approaches - sequence-based approaches like expressed sequence tags (ESTs) and serial analysis of gene expression (SAGE), and microarray-based approaches. For sequence-based approaches, it describes how ESTs can provide gene discovery and expression information but have limitations. It outlines the SAGE methodology and gene index construction to organize EST data. For microarrays, it summarizes the basic workflow including sample preparation, hybridization, image analysis and data normalization to identify differentially expressed genes through statistical tests.
TILLING & ECO-TILLING
A new era of genomics for plant science research has opened due the complete genome sequencing projects of Arabidopsis thaliana and rice. The sequence information available in public database has highlighted the need to develop genome scale reverse genetic strategies for functional analysis (Till et al., 2003). As most of the phenotypes are obscure, the forward genetics can hardly meet the demand of a high throughput and large-scale survey of gene functions. Targeting Induced Local Lesions in Genome TILLING is a general reverse genetic technique that combines chemical mutagenesis with PCR based screening to identity point mutations in regions of interest (McCallum et al., 2000). This strategy works with a mismatch-specific endonuclease to detect induced or natural DNA polymorphisms in genes of interest. A newly developed general reverse genetic strategy helps to locate an allelic series of induced point mutations in genes of interest. It allows the rapid and inexpensive detection of induced point mutations in populations of physically or chemically mutagenized individuals. To create an induced population with the use of physical/chemical mutagens is the first prerequisite for TILLING approach. Most of the plant species are compatible with this technique due to their self-fertilized nature and the seeds produced by these plants can be stored for long periods of time (Borevitz et al., 2003). The seeds are treated with mutagens and raised to harvest M1 plants, which are consequently, self-fertilized to raise the M2 population. DNA extracted from M2 plants is used in mutational screening (Colbert et al., 2001). To avoid mixing of the same mutation only one M2 plant from each M1 is used for DNA extraction (Till et al., 2007). The M3 seeds produce by selfing the M2 progeny can be well preserved for long term storage. Ethyl methane sulfonate (EMS) has been extensively used as a chemical mutagen in TILLING studies in plants to generate mutant populations, although other mutagens can be effective. EMS produces transitional mutations (G/C, A/T) by alkylating G residues which pairs with T instead of the conservative base pairing with C (Nagy et al., 2003). It is a constructive approach for users to attempt a range of chemical mutagens to assess the lethality and sterility on germinal tissue before creating large mutant populations.
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Similar to RNA interference & its application
Presentation1 Gene silencing.pptx
This seminar presentation discusses gene silencing. It introduces gene silencing as the regulation of gene expression to prevent expression of a certain gene, which can occur during transcription or translation. It then describes the main types of gene silencing - transcriptional, post-transcriptional, and meiotic gene silencing. Research methods like antisense oligonucleotides and RNA interference are explained. Applications of gene silencing in cancer, respiratory diseases, and Huntington's disease are provided. The conclusion covers that gene silencing was first observed in plants/fungi but also exists in mammals, and can be used to extinguish expression of harmful genes.
Gene silensing
Gene silencing is a process where the expression of a gene is reduced or turned off. It occurs through transcriptional or post-transcriptional mechanisms. At the transcriptional level, gene expression is modified directly. At the post-transcriptional level, double stranded RNA suppresses gene expression by binding to mRNA and preventing protein production. There are three main gene silencing mechanisms - antisense oligonucleotides, ribozymes, and RNA interference. RNAi has become the most widely used mechanism due to its effectiveness and specificity. Gene silencing has many applications including developing treatments for diseases like cancer, HIV, and respiratory illnesses. It is also used to modify crop traits and study unknown gene functions.
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This document discusses RNA interference (RNAi) and antisense RNA technology for controlling gene expression in plants. It explains that RNAi works by introducing double-stranded RNA that is processed into siRNAs to target and degrade complementary mRNA, preventing protein production. Antisense RNA also binds to mRNA but blocks processing or translation. Both techniques allow specific genes to be silenced and have applications for improving plant traits like disease resistance, ripening time, and nutrient content. The document provides detailed explanations of the mechanisms and applications of RNAi and antisense RNA technology.
Gene silencing
Gene silencing, or turning off genes, occurs naturally through epigenetic regulation and can also be induced experimentally. It was first observed in plants in the 1980s. There are three main types: transcriptional silencing by DNA/chromatin modification, post-transcriptional silencing via RNA interference, and meiotic silencing. Key components of RNAi include microRNAs, siRNAs, Dicer, and RISC. Gene silencing has applications in medical research to study diseases, in developing treatments for cancer, infectious diseases, and respiratory/neurological disorders. However, delivery issues must be addressed for therapies, as the blood-brain barrier prevents many molecules from entering the brain. Gene silencing was also used commercially to create non-b
Antisense RNA Technology Forr Crop Improvement
Antisense RNA Technology Forr Crop ImprovementJunagadh Agricultural University, Junagadh (Guj.), India
Researchers used RNA interference technology to genetically modify maize kernels for improved nutritional value. They targeted the lysine catabolism pathway by suppressing the LKR/SDH genes through recombinant RNAi. This led to transgenic maize with higher free lysine content. Experiments showed segregated reduction of LKR/SDH expression in mature F1 kernels, with consistent reduction at later developmental stages but normal accumulation earlier. Quantitative analysis confirmed LKR/SDH reduction in both embryo and endosperm, along with decreased LKR activity and increased free lysine and saccharopine levels. The study demonstrated that RNAi can effectively modify lysine metabolism in maize to enhance its nutritional profile.Rna interference
RNA interference (RNAi) is a biological process where RNA molecules inhibit gene expression by destroying mRNA. It was discovered as gene silencing in plants and nematodes. RNAi is triggered by double-stranded RNA being cut into small interfering RNAs that target homologous mRNAs for degradation. This protects against viruses and regulates genes. RNAi has many applications, including modulating HIV replication and treating cancer by silencing viral or other genes. It is a powerful new genetic tool with potential for therapeutics.
Gene silencing and rnai
Gene silencing is a method of regulating gene expression to reduce or prevent the expression of certain genes. It can occur during transcription or translation and is often used in research. Methods of gene silencing like RNAi, CRISPR, and siRNA are being researched as potential therapeutics for diseases. Gene silencing reduces gene expression by at least 70% but does not eliminate it entirely, allowing study of essential genes. RNA interference is a common method of gene silencing that uses small interfering RNAs or microRNAs to regulate gene expression.
Nucleic acids as therapeutic agents
Nucleic acids have potential as therapeutic agents by inhibiting gene expression through various mechanisms. Plasmids can introduce genetic material into cells to produce therapeutic proteins. Oligonucleotides can be used for antisense or antigene applications to selectively block expression of disease-causing proteins. Aptamers and DNAzymes can directly interact with and interfere with proteins implicated in disease. RNA-based therapeutics like RNA aptamers, decoys, antisense RNA, ribozymes, and siRNAs can also inhibit gene expression. MicroRNAs naturally downregulate gene expression and have potential therapeutic applications. Gene and stem cell therapies aim to treat genetic disorders and repair damaged tissue.
Rna interfernce
RNA interference( RNAi) is a mechanism that inhibits gene expression at post transcription level by degrading or slicing specific mRNA
Exploring Cutting-Edge Gene Silencing Approaches in Cardiovascular Disease | ...
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Discover the cutting-edge Gene Silencing approach in cardiovascular disease. Explore RNAi, ASOs, and their therapeutic applications for a healthier heart.Viral suppressors of rna silencing
This document summarizes RNA silencing and viral suppressors of RNA silencing in plants. It discusses how viruses have evolved proteins called viral suppressors of RNA silencing (VSRs) to counter the plant's RNA silencing antiviral response. It describes the mechanism of RNA silencing in plants and how various viral suppressors target different steps in the silencing pathway, such as viral RNA recognition, dicing, RISC assembly, and RNA targeting. Functional assays used to identify VSRs are also summarized, including transient expression assays, reversal of silencing assays, and stable expression assays. Finally, examples of specific viral suppressors are discussed, including tombusvirus P19 and cucumber mosaic virus protein 2b, and their mechanisms of suppressing
sirna and mirna
1. siRNA and miRNA are types of non-coding RNAs that play important roles in post-transcriptional gene regulation. siRNA induces gene silencing through RNA interference, while miRNA binds to target mRNAs to inhibit translation.
2. Both siRNA and miRNA have shown promise in developing therapies for diseases like cancer, asthma, neurodegenerative disorders, and viral infections by silencing genes involved in disease pathways. Delivery methods continue to be improved to enhance their therapeutic potential.
3. The key differences between siRNA and miRNA are their biogenesis, targeting specificity, and mechanisms of gene regulation. siRNA is synthesized from long double-stranded RNA and can perfectly complement target mRNAs, while miRNA originates from
Rna interference in insect pest management
This document discusses RNA interference (RNAi) and its potential applications for insect pest management in agriculture. It provides background on the discovery of RNAi and how it functions as a gene silencing mechanism. The document then explores how RNAi operates in insects and various methods for delivering double-stranded RNA. Both challenges and opportunities for using RNAi to control insect pests are examined, along with prospects like targeting multiple genes simultaneously or combining RNAi with other approaches like Bt toxins.
Regulatory RNA
RNA is a polymer made of ribonucleotides linked together. There are three main classes of RNA - transfer RNA, ribosomal RNA, and messenger RNA. In eukaryotes, primary transcripts undergo processing including capping, polyadenylation, and splicing before being transported to the cytoplasm for translation. MicroRNAs and small interfering RNAs are types of small regulatory RNAs that cause inhibition of gene expression through post-transcriptional gene silencing. Both miRNAs and siRNAs have potential applications as therapeutic targets in humans.
Gene knockoff
RNA interference (RNAi) is a technique that uses double-stranded RNA to silence gene expression. It involves introducing dsRNA into a cell that is complementary to the target mRNA. This dsRNA is processed by the Dicer enzyme into siRNAs which are incorporated into the RISC complex. The RISC complex uses the siRNA to identify and cleave the target mRNA, preventing it from being translated into protein. RNAi provides a valuable tool for studying gene function and has applications in biotechnology, agriculture, and medicine by allowing researchers to knock down gene expression.
Viral suppressors of rna silencing
RNA silencing is an ancient eukaryotic pathway in which double stranded RNA (dsRNA) triggers destruction of related RNAs in the cell.This study focuses on the currently known plant virus encoded suppressors of silencing and the functional assays used to identify these proteins. Because they interfere with silencing at different points in the pathway, these viral suppressors are powerful tools to help unravel the mechanism of RNA silencing in plants.
Gene silencing
Gene silencing is a technique that reduces or eliminates protein production from a gene. It occurs through RNA interference, where small interfering RNAs are processed by an enzyme called Dicer and loaded into an RNA-induced silencing complex (RISC) that targets complementary mRNAs for degradation. There are two main types of gene silencing - transcriptional which alters DNA accessibility, and post-transcriptional via RNAi technologies. RNAi has therapeutic applications for cancer, infectious diseases, and neurodegenerative disorders by knocking down target genes. The first approved RNAi drug, Patisiran, treats hereditary transthyretin-mediated amyloidosis.
Rna interfernce ppt
RNA interference (RNAi) is a mechanism where double-stranded RNA inhibits gene expression. It was discovered in plants, fungi, and animals. The mechanism involves dicer enzymes cleaving long double-stranded RNA into short interfering RNAs (siRNAs). These siRNAs are incorporated into an RNA-induced silencing complex (RISC) which guides the complex to mRNAs with complementary sequences, resulting in their degradation. RNAi has applications in therapeutics for cancer, viruses, and genetic disorders, as well as research in gene function and pathways.
Gene Silencing
This document discusses gene silencing, which is the suppression or interruption of gene expression at the transcriptional or translational level. It regulates gene expression and prevents certain genes from being expressed. There are several types of gene silencing, including transcriptional silencing through genomic imprinting, paramutation, and transposon silencing. Post-transcriptional gene silencing occurs through RNA interference pathways using small interfering RNA, microRNA, Dicer, and RISC complex. Research methods for gene silencing include antisense oligonucleotides, ribozymes, siRNA, and microRNA. Gene silencing has applications in biotechnology and medicine, with advantages such as being cost effective and having high specificity, though it also has disadvantages like potential toxicity.
EPIGENETIC.pptx
Gene silencing is a technique that reduces or eliminates the production of a protein from its corresponding gene through mechanisms other than genetic modification. There are two main types of gene silencing - transcriptional and post-transcriptional. Post-transcriptional gene silencing includes RNA interference (RNAi), which involves introducing double-stranded RNA that binds to mRNA and prevents replication, effectively silencing gene expression in a sequence-specific manner. RNAi is a powerful tool for gene function analysis and applications in biotechnology and agriculture.
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RNA interference & its application
- 1. R N A I N T E R F E R E N C E A N D I T S A P P L I C AT I O N S B Y - A K R I T I G A U R A V
- 2. Overview 1. Introduction 2. Mechanism 3. Recent Application 4. Conclusion
- 3. Several terms are used to described RNA silencing; usually there are three phenotypically different but mechanistically similar phenomena: 1. Co-suppression or post-transcriptional gene silencing (PTGS) in plants 2. Quelling in fungi 3. RNA interference in animal kingdom Introduction
- 4. Definition RNA interference (RNAi) is a mechanism that inhibits gene expression at the stage of translation or by hindering the transcription of specific genes.
- 6. Recent applications of RNAi • Modulation of HIV-1 replication by RNA interference. • Potent and specific inhibition of human immunodeficiency virus type 1 replication by RNA interference. • In the treatment of Hepatitis. • It is used to intervene in the process of atherosclerosis or to reduce damage to heart tissue and brain cells.
- 7. • RNAi is a powerful and attractive genetic approach because of the diversity of its applications. The potential uses currently in progress include the identification of specific gene functions in living systems and creation of genome wide screens. Development of antiviral and anticancer therapies are broadening the horizons of the therapeutic arena. • Another value of RNAi screens is in combining it with other functional genomic assays enabling mapping of biochemical pathways. Impact of RNAi is also being extended to the field of agriculture for example by increasing disease resistance in plants. Conclusions