CMMB 403 Lecture Notes - Lecture 29: Guide Rna, Cas9, Crispr
6 Jan 2019
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To understand the function of a specific gene during development, we usually want to inactivate the gene and see how development proceeds in its absence reverse genetics. Reverse genetics: know there is a gene, inactivate it, see phenotype, infer function of gene. Things we"ve been able to do only in mice till now, we can now do it in other organism. Forward genetic screen: phenotype is first generated, and then the gene responsible is cloned/characterized later. Compared to earlier methods, crispr/cas9 is a much simpler and faster method to introduce changes into genomes. Crispr/cas9 uses a nuclease and a 20 nucleotide rna sequence to target specific sites in the genome, which can easily be exploited to study gene function during development. Nuclease which is used to cleave dna is cas9 cleaves at only specific point in genome using guide rna which has 20 nucleotide sequence that base pairs with a complementary sequence in the genome.
