The human CRISPR ′Brunello′ pooled library is designed using optimized metrics, as published by, Doench et al. Nat Biotechnol. (2016) and described further in Sanson, K.R., et al. Nat Commun (2018), which combine improved on-target activity predictions (Rule Set 2) with an off-target score, the Cutting Frequency Determination (CFD). The library is designed to be compact and efficient to maximize screening efficiency and performance.
Custom pools for follow-up screening or 10x Genomics Compatible CRISPR pools are also available by contacting your local sales representative.
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.