Advanced CRISPR Cas9 Genetic Engineering
The CRISPR Cas9 system has been harnessed to create a simple, RNA programmable method to mediate genome editing in mammalian cells, and can be used to generate gene knockouts (via insertion/deletion) or knock-ins via homology directed repairs (HDR). And to create gene disruptions resulting in permanent mutations, a single guide RNA (sgRNA) is generated to direct the Cas9 nuclease to a specific genomic location. Cas9-induced double strand breaks are repaired via the non-homologous end joining (NHEJ) DNA repair pathway. The repair is error prone, and thus insertions and deletions (INDELs) may be introduced that can disrupt gene function. CRISPR Cas9 technology has revolutionized genome editing, allowing a previously unattainable level of genomic targeting, efficiency, and simplicity.