酵母基因编辑技术从单基因操作到基因组重建-遗传.PDF

Hereditas (Beijing) 2015 年 10 月, 37(10): 1021―1028 综 述 酵母基因编辑技术：从单基因操作到基因组重建 李晨，楼慧强 中国农业大学生物学院，农业生物技术国家重点实验室，北京 100193 摘要:DNA 双链发生断裂(Double strand break, DSB) 时，细胞通常会采用同源重组(Homologous recombination, HR)这一主要修复方式。基于这一原理，人们开发出多种技术引入双链断裂从而实现对基因组 DNA 序列的靶 向编辑。真核生物基因敲除或编辑最早通过在外源片段中引入双链断裂和同源臂在酵母中获得成功。30 多年 来，酵母遗传操作走过了一条从单基因单位点到多基因多位点，乃至最新的基因组合成与重建之路。本文沿着 引入DNA 双链断裂技术的发展与演变这一主线，对一系列基因编辑及基因组编辑技术的发展进行了简要回顾 和综述，旨在理清基因编辑技术的发展脉络，也为高等哺乳动物的基因编辑技术体系的建立和完善提供参考。 DNA 双链断裂；基因编辑；基因组编辑；基因组合成与重建；酵母 关键词: From gene editing to genome reconstitution: evolving techniques in yeast Chen Li, Huiqiang Lou State Key Laboratory of Agro-Biotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China Abstract: Homologous recombination is one of the main repair pathways in response to DNA double strand break (DSB) in eukaryotes. Based on this, a series of techniques to introduce DSB have been developed in order to edit the DNA sequence of genome. In eukaryotes, the gene editing technique was first established in S . cerevisiae by trans- formation of a foreign DNA fragment containing the sequence homologous to the targeted site more than thirty years ago. The core of all currently available editing methods lies in the introduction of DSB. Here, we try to convey a historic view of various editing techniques from its original version to the up-to-dated genome synthesis and recons- titution. We believe that this review will help to illustrate the trend of the development of genome editing techniques, which will provide a valuable reference for developing similar techniques in mammals. Keywords: DNA double strand break; gene editing; genome editing; genome synthesis/reconstitution; yeast 同源重组(Homologous recombination)是指发生 了减数分裂等过程中程序性发生的同源重组，这一 在