CRISPR knockout screening outperforms shRNA and CRISPRi in identifying essential genes
CRISPR knockout screening outperforms shRNA and CRISPRi in identifying essential genes
复制标题
DOI：
10.1038/nbt.3536
复制
发表时间：
2016-06-01
影响因子：
46.9
通讯作者：
Bernards, Rene
中科院分区：
文献类型：
--
作者：
Evers, Bastiaan;Jastrzebski, Katarzyna;Bernards, Rene
高通量基因筛选已成为研究各种生物过程的重要工具。在这里，我们通过实验比较了基于成簇的规则间隔短回文重复序列（CRISPR）/CRISPR相关蛋白9（Cas9）或其转录抑制变体CRISPR干扰（CRISPRi）的系统，以及传统的基于短发夹RNA（shRNA）的系统进行致死性筛选。我们发现CRISPR技术表现最好，噪音低，脱靶效应最小，试剂之间的活性一致。
High-throughput genetic screens have become essential tools for studying a wide variety of biological processes. Here we experimentally compare systems based on clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) or its transcriptionally repressive variant, CRISPR-interference (CRISPRi), with a traditional short hairpin RNA (shRNA)-based system for performing lethality screens. We find that the CRISPR technology performed best, with low noise, minimal off-target effects and consistent activity across reagents.