Improved vectors and genome-wide libraries for CRISPR screening

作者：Neville E. Sanjana, Ophir Shalem, Feng Zhang · 发表于：Nature Methods · 年份：2014 · DOI：10.1038/nmeth.3047 · 被引用次数：4959 · 研究领域：Biology、Medicine

Genome-wide, targeted loss-of-function pooled screens using the CRISPR (clustered regularly interspaced short palindrome repeats)–associated nuclease Cas9 in human and mouse cells provide an alternative screening system to RNA interference (RNAi) and have been used to reveal new mechanisms in diverse biological models1–4. Previously, we used a Genome-scale CRISPR Knock-Out (GeCKO) library to identify loss-of-function mutations conferring vemurafenib resistance in a melanoma model1. However, initial lentiviral delivery systems for CRISPR screening had low viral titer or required a cell line already expressing Cas9, limiting the range of biological systems amenable to screening.