Towards a toolbox for mammalian gene editing: Removing off-target activity and protein interactions of Cas9

在过去的30年里，分子遗传学研究在哺乳动物基因组测序和注释方面取得了前所未有的突破。研究人员目前面临的主要挑战之一是如何有效地为每个基因分配功能。虽然研究人员通常分析与基因功能丧失相关的表型变化，但在哺乳动物生物体中执行基因敲除的经典技术是缓慢、费力和昂贵的。最近，DNA序列特异性核酸酶(SSN)，如TALENS和CRISPR/CAS9，已经成为有望使这一过程发生革命性变化的工具。我的实验室的一个总体目标是开发强大的技术，以高精度地从基因上改变哺乳动物生物体。通过结合分子进化和蛋白质组学的方法，该计划将为哺乳动物开发一个实现这一可能性的基因编辑工具箱。

In the past 30 years, molecular genetics research has resulted in unprecedented breakthroughs in sequencing and annotating the mammalian genome. One of the main challenges now facing researchers is how to efficiently assign function to each gene. While researchers typically analyze phenotypic changes associated with gene loss-of-function, classical techniques for performing gene knockout in mammalian organisms are slow, laborious, and costly. Recently, DNA sequence-specific nucleases (SSNs) such as TALENs and CRISPR/Cas9 have emerged as tools that are poised to revolutionize this process. An over-arching goal of my lab is to develop robust technology for genetically altering mammalian organisms with high precision. Through a combination of molecular evolution and proteomics approaches, this program will develop a gene editing toolbox for mammals that will realize this possibility.