GENE 320 Lecture Notes - Lecture 23: Homology Directed Repair, Crispr, Nuclease
Document Summary
Genome size in eukaryotes is highly variable because of The human genome has about 20,000 genes and an estimated 100,000 proteins. Comparative genomics studies several model organisms, whose genomes revealed interesting elements of the human genome. Interconnected through genome evolution, speciation, and gene expression. To manipulate speci c allele, locus, or base sequence and learn its function on gene of interest. To disrupt or eliminate speci c gene/genes and see what happens. Creating knockout mice create segment of dna for introduction into cell undergoes homologous recombination with gene of interest and renders it nonfunctional. Target vector has mutated copy of gene of interest. Foregin dna disrupts reading frame and produces nonfunctional protein. Es cell takes in targeting vector and endogenous enzyme recombinase catalyzes homologous recombination. Recombinant es cells are injected into mouse embryo. Allows scientist to control when target gene is disrupted. Nuclease induced double strand breaks can be repaired by nonhomologous end joining or homology directed repair pathways.
