CRISPR/Cas9技术存在的问题及其改进措施的研究进展

Current Issues and Progress in the Application of CRISPR/Cas9 Technique

CRISPR/Cas9技术是一种RNA引导的基因组编辑技术,能对基因进行精确性敲除、敲入、替换等,从而实现探究基因功能、修复致病基因等目的。该技术凭借操作简便、价格低廉、可对基因位点进行编辑、可拓展性强等优势,在近几年迅速发展完善,已成为继锌指核酸酶技术和类转录激活因子效应物核酸酶技术之后的第三大基因组编辑技术。在简单介绍CRISPR/Cas9技术的作用机制后,主要针对该技术现阶段面临的精确修复比例低、PAM限制识别序列、脱靶现象、马赛克现象的问题以及相应的改进措施进行阐述,旨在让研究者客观的认识CRISPR/Cas9技术存在的不足之处,为合理使用或改进该技术提供系统的文献综述。

CRISPR/Cas9[clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9]is a RNA-guidedgenome-editing technique,which can be applied in the deletion,insertion or replacement of specific DNA sequence thus to verify the functionof genes or remedy pathogenic genes. Owing to its simplicity,affordability,multiple-site editing and scalability,the technique of CRISPR/Cas9 is undergoing rapid development and perfection and has been one of the first three prevailing genome-editing technology following thatof Zinc finger nucleases and transcription activator-like effector nucleases. In this review,we focus on the current issues of CRISPR/Cas9,including low efficiency of precise-editing,sequence-recognizing limitation from PAM,off-target,unavoidable mosaicism,together withcorresponding tactics for improvement. Our aim is to provide the objective understanding on the current disadvantage of CRISPR/Cas9 techniqueto researchers and the systematic review for reasonable application and improvement of this technology.