CRISPR/Cas9基因编辑在三维基因组研究中的应用  

Probing 3D genome by CRISPR/Cas9

作　　者：刘沛峰 吴强 Peifeng Liu;Qiang Wu(Center for Comparative Biomedicine,Key Laboratory of Systems Biomedicine(Ministry of Education),Institute of Systems Biomedicine,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学系统生物医学研究院比较生物医学研究中心系统生物医学教育部重点实验室

出　　处：《遗传》2020年第1期18-31,共14页Hereditas（Beijing)

基　　金：国家自然科学基金项目(编号：31630039,91640118);国家重点研发计划课题项目(编号：2017YFA0504203,2018YFC1004504);上海市科学技术委员会项目(编号：19JC1412500)资助~~

摘　　要：CRISPR/Cas9系统在基因编辑方面具有巨大优势,能够低成本、可编程、方便快捷地用于动物、植物以及微生物的基因组靶向编辑和功能改造。三维基因组学是近年来兴起的一门研究染色质高级结构动态调控及基因组生物学功能的交叉学科。在三维基因组研究中,通常采用对DNA片段进行基因编辑以模拟基因组结构性变异,标记特定DNA片段,进而研究调控元件对于基因调控、细胞分化、组织发生、器官形成、个体发育的影响,最终阐明三维基因组的组装调控机制和生物学功能。因此,CRISPR及其衍生技术为研究三维基因组提供了极好的遗传学工具。本文主要综述了CRISPR片段编辑及其衍生技术在三维基因组调控与功能研究中的应用,以期为后续研究工作提供理论参考以及新的研究思路。CRISPR/Cas9 system has significant advantages in gene editing strategy,offering cost-effective and efficient means to modify and edit the genomes of animals,plants,and microorganisms.Three-dimensional(3D)genome is an emerging and interdisciplinary field catapulted by combined technological breakthroughs of chromosome conformation capture with next-generation sequencing and live imaging with super-resolution microscopy.An important aspect of 3D genomics is to model structural variations and label specific genomic fragments to investigate the effects of manipulation of genomic elements on gene expression regulation,cell development and differentiation,and spatial location of chromosomal regions.Therefore,CRISPR/Cas9 system and its derivative technologies of DNA-fragment editing are excellent toolboxes for investigating dynamics and functions of the higher-order chromatin organization and three-dimensional genome structure.In this review,we describe the opportunities and challenges of CRISPR as well as its derivative technologies in 3D genome research,thereby providing some critical references and future research directions in the field.

关 键 词：CRISPR/Cas9系统 三维基因组 DNA片段编辑 染色质重排 Cas9核酸酶内切机制 

分 类 号：Q78[生物学—分子生物学]