CMMB 461 Lecture Notes - Lecture 20: Homologous Recombination, Cas9, Crispr

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Double stranded break in dna (made by crispr) will be repaired by two pathways. Non-homologous end joining (nhej) which is error prone and introduces deletion and insertion (indel mutations) Way of disrupting function of gene, making it a gof mutation by deleting negative regulator in promoter, so gene expressed even more. This pathway is error prone, because when it connects two strands, you could get some nucleotides that are either deleted or added. Homologous recombination uses the wt copy of the chromosome or donor dna for gene correction. This is basically cutting and rejoining dna perfectly. In this event, there is a perfect exchange of dna from homologous chromosomes. Potentially get a wt copy of chromosome or get donor dna. Machinery uses other chromosome as a template. If you have a defective gene that you want to fix, you can use this technique to revert mutated gene back into wt copy of the gene.

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