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体细胞CRISPR-Cas9在秀丽隐杆线虫条件性基因组编辑中的应用。

The application of somatic CRISPR-Cas9 to conditional genome editing in Caenorhabditis elegans.

作者信息

Li Wei, Ou Guangshuo

机构信息

School of Medicine, Tsinghua University, Beijing, 100084, China.

Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, 100084, China.

出版信息

Genesis. 2016 Apr;54(4):170-81. doi: 10.1002/dvg.22932. Epub 2016 Apr 14.

Abstract

Forward and reverse genetic approaches have been well developed in the nematode Caenorhabditis elegans; however, efficient genetic tools to generate conditional gene mutations are still in high demand. Recently, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR-Cas9) system for genome modification has provided an additional tool for C. elegans researchers to achieve simple and efficient conditional targeted mutagenesis. Here, we review recent advances in the somatic expression of Cas9 endonuclease for conditional gene editing. We present some practical considerations for improving the efficiency and reducing the off-target effects of somatic CRISPR-Cas9 and highlight a strategy to analyze somatic mutation at single-cell resolution. Finally, we outline future applications and consider challenges for this emerging genome editing platform that will need to be addressed in the future.

摘要

正向和反向遗传学方法在线虫秀丽隐杆线虫中已经得到了很好的发展;然而,对于生成条件性基因突变的高效遗传工具的需求仍然很高。最近,用于基因组修饰的成簇规律间隔短回文重复序列/CRISPR相关蛋白9(CRISPR-Cas9)系统为秀丽隐杆线虫研究人员提供了另一种工具,以实现简单高效的条件性靶向诱变。在这里,我们回顾了用于条件性基因编辑的Cas9核酸内切酶体细胞表达的最新进展。我们提出了一些提高体细胞CRISPR-Cas9效率和减少脱靶效应的实际考虑因素,并强调了一种在单细胞分辨率下分析体细胞突变的策略。最后,我们概述了该新兴基因组编辑平台的未来应用,并考虑了未来需要解决的挑战。

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