一般描述
The Human Whole Genome Toronto KnockOut Library v3 (TKOv3) is optimized for editing efficiency using empirical data described in the findings by the Moffat lab.The compact library contains 70,948 gRNAs targeting 18,053 protein-coding genes (4 gRNAs/gene) with 142 control gRNAs targeting EGFP, LacZ, and luciferase (71,090 total). The optimized library size of TKOv3 offers improved accuracy, efficiency, and scalability for CRISPR KO screens.
Custom pools for follow-up screening or 10x Genomics Compatible CRISPR pools are also available by contacting your local sales representative.
原理
In a CRISPR KO screen, Cas9 introduces double-strand breaks at locations specified by a gRNA. When the endogenous non-homologous end-joining (NHEJ) DNA repair system corrects these breaks, this often leads to the introduction of frame-shift mutations that effectively knock out the gene. Thus, the power of CRISPR for genome engineering, coupled with the ability to perform large-scale, whole-genome loss-of-function (LOF) screening, has allowed breakthroughs in identifying gene pathways in drug resistance and disease.