基因编辑阳性细胞的富集报告系统

引用本文: 张力弦, 王启扉, 曹杨, 叶承宇, 陈智洋, 徐启杰, 邹秉杰, 宋沁馨, 周国华. 基因编辑阳性细胞的富集报告系统[J]. 分析化学, 2020, 48(1): 1-12. doi: 10.19756/j.issn.0253-3820.191322 shu

Citation: ZHANG Li-Xian, WANG Qi-Fei, CAO Yang, YE Cheng-Yu, CHEN Zhi-Yang, XU Qi-Jie, ZOU Bing-Jie, SONG Qin-Xin, ZHOU Guo-Hua. Enrichment Reporter System of Genome Editing Positive Cells[J]. Chinese Journal of Analytical Chemistry, 2020, 48(1): 1-12. doi: 10.19756/j.issn.0253-3820.191322 shu

基因编辑阳性细胞的富集报告系统

1.
药物质量与安全预警教育部重点实验室, 中国药科大学, 南京 210009;
2.
东部战区总医院药理科, 南京 210002
基金项目:
本文系国家自然科学基金项目（No.81673390）、江苏省基础研究计划自然科学基金项目（No.BK20191322）、江苏省杰出青年基金项目（No.BK20180005）、药物质量与安全预警教育部重点实验室项目（No.DQCP2017MS01）、中国药科大学药学基地科研训练及科研能力提高项目（No.J1310032）和江苏省青蓝工程项目资助

摘要: 基因编辑技术已成为医学生物学研究的重要工具，将不同基因快速高效地富集到基因编辑阳性细胞并进行针对性研究是促进基因编辑技术发展的关键。近年来，基于非同源末端接合、同源直接修复、单链退火、倒位重排等基因组修复机制，研究者利用荧光蛋白或抗性标签基因修复后表达的原理，建立了多种可用于基因编辑阳性细胞筛选与富集的报告系统。富集后的细胞采用T7E1酶切法或测序技术进行突变分析，呈现显著降低的背景信号和增加的突变比例。此类报告系统有利于基因编辑效果的表征。此外，分选后的阳性细胞可继续培养，在突变细胞系构建及突变细胞功能研究等领域具有良好的发展前景。本文对这些报告系统的设计原理与应用进行了总结，以期为建立更加完善的基因编辑评价系统提供参考。

关键词:

English

Enrichment Reporter System of Genome Editing Positive Cells

1.
Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing 210009, China;
2.
Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China
Received Date: 06 June 2019
Revised Date: 08 November 2019
Fund Project:
This work was supported by the National Natural Science Foundation of China (No.81673390), the Jiangsu Provincial Natural Science Foundation (No. BK20191322), the Jiangsu Provincial Science Fund for Distinguished Young Scholars (No.BK20180005), the Open Project Program of MOE Key Laboratory of Drug Quality Control and Pharmacovigilance (No. DQCP2017MS01), the College Students Innovation Project for the R&D of Novel Drugs (No.J1310032), and the Qing Lan Project of Jiangsu Province.

Abstract: Genome editing has become a vital tool in medical biology research. The critical mission of facilitating the progress of genome editing is to enrich positive edited cells quickly and effectively. In recent years, researchers have established various reporter systems for selection and enrichment of editing-induced positive cells based on genome repair mechanisms, such as non-homologous end joining, homology directed repair, single strand annealing and inversion, and the principle of the expression of fluorescent protein or resistance tag after genome repair. The T7E1 assay or sequencing method can analyze the mutation of enriched cells with lower background signals and higher mutation ratio. Therefore, these reporter systems can profit the characterization of genome editing effectiveness. Besides, positive cells can be cultured continuously, so this technology possesses a promising prospect of mutated cell line construction and the research of mutated cell functions. This article summarized the design principles and applications of these reporter systems and provided a reference to construct a more perfect evaluating system for genome editing.

Key words:

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

基因编辑阳性细胞的富集报告系统

English

Enrichment Reporter System of Genome Editing Positive Cells

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基因编辑阳性细胞的富集报告系统

English

Enrichment Reporter System of Genome Editing Positive Cells

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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