基因编辑阳性细胞的富集报告系统

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

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

  • 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。

English


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  • 收稿日期:  2019-06-06
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