Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy

被引:152
作者
Liu, Jiyong [2 ]
Li, Changqing [2 ]
Yu, Zhongsheng [2 ]
Huang, Peng [3 ]
Wu, Honggang [2 ]
Wei, Chuanxian [2 ]
Zhu, Nannan [2 ]
Shen, Yan [3 ]
Chen, Yixu [2 ]
Zhang, Bo [3 ]
Deng, Wu-Min [1 ]
Jiao, Renjie [2 ]
机构
[1] Florida State Univ, Dept Biol Sci, Tallahassee, FL 32304 USA
[2] Chinese Acad Sci, Inst Biophys, State Key Lab Brain & Cognit Sci, Beijing 100101, Peoples R China
[3] Peking Univ, Coll Life Sci, Minist Educ, Key Lab Cell Proliferat & Differentiat, Beijing 100871, Peoples R China
基金
美国国家科学基金会;
关键词
TALEN; Drosophila; Genomic modification; Unit assembly; Reverse genetics; ZINC-FINGER NUCLEASES; ARTIFICIAL TRANSCRIPTION FACTORS; HOMOLOGOUS RECOMBINATION; DNA-SEQUENCES; III EFFECTORS; RECOGNITION; CONSTRUCTION; DOMAINS; FAMILY; MUTAGENESIS;
D O I
10.1016/j.jgg.2012.04.003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Technology development has always been one of the forces driving breakthroughs in biomedical research. Since the time of Thomas Morgan, Drosophilists have, step by step, developed powerful genetic tools for manipulating and functionally dissecting the Drosophila genome, but room for improving these technologies and developing new techniques is still large, especially today as biologists start to study systematically the functional genomics of different model organisms, including humans, in a high-throughput manner. Here, we report, for the first time in Drosophila, a rapid, easy, and highly specific method for modifying the Drosophila genome at a very high efficiency by means of an improved transcription activator-like effector nuclease (TALEN) strategy. We took advantage of the very recently developed "unit assembly" strategy to assemble two pairs of specific TALENs designed to modify the yellow gene (on the sex chromosome) and a novel autosomal gene. The mRNAs of TALENs were subsequently injected into Drosophila embryos. From 31.2% of the injected F-0 fertile flies, we detected inheritable modification involving the yellow gene. The entire process from construction of specific TALENs to detection of inheritable modifications can be accomplished within one month. The potential applications of this TALEN-mediated genome modification method in Drosophila are discussed.
引用
收藏
页码:209 / 215
页数:7
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