基于CRISPR/Cas9的基因分析方法研究进展

自2012年首次证明了CRISPR/Cas9可以在体外进行DNA切割试验以来,CRISPR技术逐渐在基因编辑研究中获得了迅速的发展,除了应用于基因编辑领域之外,它在基因表达调控、基因成像、基因分析等方面也展现出了巨大的应用潜力。尤其在基因分析领域,CRISPR技术由于其精确的基因识别、室温的反应条件、易设计性和操作性等特色,使得一系列新型的基因检测技术得以发展,并取得了超越常规技术的一些检测参数。本文以Cas9蛋白为对象,综述了近些年来在该领域取得的研究进展。主要论述Cas9蛋白的功能、改造、引导RNA(sgRNA)的设计及其在基因分析方法上的应用。

Research Progress of Genetic Analysis Methods Based on CRISPR/Cas9

Since the first demonstration of in vitro CRISPR/Cas9 DNA cleavage experiment in 2012,genetic editing research based on CRISPR technology has been greatly developed.In addition to its application in the field of gene editing,CRISPR technology has also shown great potential in gene expression regulation,gene imaging,and gene analysis.Especially in the field of genetic analysis,CRISPR technology has been developed for constructing a series of new gene detection technologies,due to its precise gene recognition,room temperature reaction conditions,ease of design and operability.CRISPR based gene analysis methods have obtained some detection parameters and gained an advantage over conventional techniques.In this paper,the research progress in this field based on Cas9 in recent years is reviewed.This paper mainly discusses the development and progress of genetic analysis methods based on the function of Cas9,engineering of guided RNA(sgRNA)and Cas9 protein.