Firstly, this how CRISPR works : the cas9 enzyme acts as the scissors and the single guided RNA (sgRNA) acts as the The native Cas9-mediated genome is completed with two steps. In the begining, Cas9 induces a DSB at on one targeted site on the genomic DNA which is guided by a 20-nt guide sequence in the crRNA. Next, the DSBs either undergo HR or NHEJ pathway.
Cas9 can be utilized to ease a broad diversity of targeted genome engineering applications. Using traditional manipulation genetic techniques, the Cas9 nuclease has enabled efficient and targeted genetic manipulation strategies. CRISPRs ease of simply designing a short RNA sequence to retarget Cas9 enables a large- scale of unbiased genome perturbation experiments to elucidate cause genetic variants or to probe gene function.
Cas9-mediated genome editing has allowed rapid generation of transgenic model and widens biological research over classic, genetically tractable animal model organisms (Sander, Joung, 2014).