sgRNA 工程提高基因组编辑和扩展功能测定。

SgRNA engineering for improved genome editing and expanded functional assays.

机构信息

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou 310027, China.

出版信息

Curr Opin Biotechnol. 2022 Jun;75:102697. doi: 10.1016/j.copbio.2022.102697. Epub 2022 Feb 23.

DOI:10.1016/j.copbio.2022.102697

PMID:35217295

Abstract

The CRISPR/Cas system has been established as the most powerful and practical genome engineering tool for both fundamental researches and biotechnological applications. Great efforts have been devoted to engineering the CRISPR system with better performance and novel functions. As an essential component, single guide RNAs (sgRNAs) have been extensively designed and engineered with desirable functions. This review highlights representative studies that optimize the sgRNA nucleotide sequences for improved genome editing performance (e.g. activity and specificity) as well as add extra aptamers and end extensions for expanded CRISPR-based functional assays (e.g. transcriptional regulation, genome imaging, and prime editor). The perspectives for further sgRNA engineering to establish more powerful and versatile CRISPR/Cas systems are also discussed.

摘要

CRISPR/Cas 系统已被确立为基础研究和生物技术应用中最强大和实用的基因组工程工具。人们已经付出了巨大的努力来对 CRISPR 系统进行工程改造，以获得更好的性能和新的功能。作为一个重要组成部分，单引导 RNA（sgRNA）已经被广泛设计和工程化，以具有理想的功能。这篇综述重点介绍了代表性的研究，这些研究优化了 sgRNA 核苷酸序列，以提高基因组编辑性能（例如活性和特异性），并添加额外的适体和末端延伸，以扩展基于 CRISPR 的功能测定（例如转录调控、基因组成像和 Prime 编辑）。还讨论了进一步 sgRNA 工程以建立更强大和多功能的 CRISPR/Cas 系统的观点。