title: Efficient and specific modifications of the Drosophila genome by means of an easy TALEN strategy.
abstract: 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.
authors.affiliation: State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, The Chinese Academy of Sciences, Datun Road 15, Beijing 100101, China.
authors.forename: Jiyong
authors.initials: J
authors.lastname: Liu
authors.forename: Changqing
authors.initials: C
authors.lastname: Li
authors.forename: Zhongsheng
authors.initials: Z
authors.lastname: Yu
authors.forename: Peng
authors.initials: P
authors.lastname: Huang
authors.forename: Honggang
authors.initials: H
authors.lastname: Wu
authors.forename: Chuanxian
authors.initials: C
authors.lastname: Wei
authors.forename: Nannan
authors.initials: N
authors.lastname: Zhu
authors.forename: Yan
authors.initials: Y
authors.lastname: Shen
authors.forename: Yixu
authors.initials: Y
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.forename: Wu-Min
authors.initials: WM
authors.lastname: Deng
authors.forename: Renjie
authors.initials: R
authors.lastname: Jiao
mesh_terms: D000818:Animals; D001483:Base Sequence; D003851:Deoxyribonucleases / Q000378:metabolism; D004330:Drosophila / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D005123:Eye / Q000254:growth & development / Q000378:metabolism; D005260:Female; D005821:Genetic Techniques / Q000295:instrumentation; D049750:Genome, Insect; D008297:Male; D008969:Molecular Sequence Data
chemical_list: D029721:Drosophila Proteins; D003851:Deoxyribonucleases