加拿大多伦多大学Jason Moffat、Benjamin J. Blencowe、Thomas Gonatopoulos-Pournatzis等研究人员合作取得新进展。他们利用Cas9–Cas12a杂交平台实现对遗传相互作用的绘制与外显子功能基因组学的解析。这一研究成果于2020年3月16日在线发表在《自然—生物技术》杂志上。
Title: Genetic interaction mapping and exon-resolution functional genomics with a hybrid Cas9–Cas12a platform
Author: Thomas Gonatopoulos-Pournatzis, Michael Aregger, Kevin R. Brown, Shaghayegh Farhangmehr, Ulrich Braunschweig, Henry N. Ward, Kevin C. H. Ha, Alexander Weiss, Maximilian Billmann, Tanja Durbic, Chad L. Myers, Benjamin J. Blencowe, Jason Moffat
Abstract: Systematic mapping of genetic interactions (GIs) and interrogation of the functions of sizable genomic segments in mammalian cells represent important goals of biomedical research. To advance these goals, we present a CRISPR (clustered regularly interspaced short palindromic repeats)-based screening system for combinatorial genetic manipulation that employs coexpression of CRISPR-associated nucleases 9 and 12a (Cas9 and Cas12a) and machine-learning-optimized libraries of hybrid Cas9–Cas12a guide RNAs. This system, named Cas Hybrid for Multiplexed Editing and screening Applications (CHyMErA), outperforms genetic screens using Cas9 or Cas12a editing alone. Application of CHyMErA to the ablation of mammalian paralog gene pairs reveals extensive GIs and uncovers phenotypes normally masked by functional redundancy. Application of CHyMErA in a chemogenetic interaction screen identifies genes that impact cell growth in response to mTOR pathway inhibition. Moreover, by systematically targeting thousands of alternative splicing events, CHyMErA identifies exons underlying human cell line fitness. CHyMErA thus represents an effective screening approach for GI mapping and the functional analysis of sizable genomic regions, such as alternative exons.