Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Nat Methods. 2016 Jan;13(1):41-50. doi: 10.1038/nmeth.3684.
The simplicity of site-specific genome targeting by type II clustered, regularly interspaced, short palindromic repeat (CRISPR)-Cas9 nucleases, along with their robust activity profile, has changed the landscape of genome editing. These favorable properties have made the CRISPR-Cas9 system the technology of choice for sequence-specific modifications in vertebrate systems. For many applications, whether the focus is on basic science investigations or therapeutic efficacy, activity and precision are important considerations when one is choosing a nuclease platform, target site and delivery method. Here we review recent methods for increasing the activity and accuracy of Cas9 and assessing the extent of off-target cleavage events.
由 II 型簇状、规律间隔、短回文重复（CRISPR）-Cas9 核酸酶介导的靶向特定基因的方法具有简单性，再加上其强大的活性特征，改变了基因组编辑的格局。这些有利的特性使得 CRISPR-Cas9 系统成为脊椎动物系统中序列特异性修饰的首选技术。对于许多应用，无论是关注基础科学研究还是治疗效果，在选择核酸酶平台、靶位点和递送方法时，活性和精度都是重要的考虑因素。在这里，我们综述了提高 Cas9 活性和准确性的最新方法，并评估了脱靶切割事件的程度。
