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
相关论文
共 42 条
  • [1] Efficient gene targeting in Drosophila with zinc-finger nucleases
    Beumer, K
    Bhattacharyya, G
    Bibikova, M
    Trautman, JK
    Carroll, D
    [J]. GENETICS, 2006, 172 (04) : 2391 - 2403
  • [2] Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases
    Beumer, Kelly J.
    Trautman, Jonathan K.
    Bozas, Ana
    Liu, Ji-Long
    Rutter, Jared
    Gall, Joseph G.
    Carroll, Dana
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (50) : 19821 - 19826
  • [3] Bibikova M, 2002, GENETICS, V161, P1169
  • [4] An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases
    Bischof, Johannes
    Maeda, Robert K.
    Hediger, Monika
    Karch, Francois
    Basler, Konrad
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (09) : 3312 - 3317
  • [5] Xanthomonas AvrBs3 Family-Type III Effectors: Discovery and Function
    Boch, Jens
    Bonas, Ulla
    [J]. ANNUAL REVIEW OF PHYTOPATHOLOGY, VOL 48, 2010, 48 : 419 - 436
  • [6] Breaking the Code of DNA Binding Specificity of TAL-Type III Effectors
    Boch, Jens
    Scholze, Heidi
    Schornack, Sebastian
    Landgraf, Angelika
    Hahn, Simone
    Kay, Sabine
    Lahaye, Thomas
    Nickstadt, Anja
    Bonas, Ulla
    [J]. SCIENCE, 2009, 326 (5959) : 1509 - 1512
  • [7] TAL Effectors: Customizable Proteins for DNA Targeting
    Bogdanove, Adam J.
    Voytas, Daniel F.
    [J]. SCIENCE, 2011, 333 (6051) : 1843 - 1846
  • [8] Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting
    Cermak, Tomas
    Doyle, Erin L.
    Christian, Michelle
    Wang, Li
    Zhang, Yong
    Schmidt, Clarice
    Baller, Joshua A.
    Somia, Nikunj V.
    Bogdanove, Adam J.
    Voytas, Daniel F.
    [J]. NUCLEIC ACIDS RESEARCH, 2011, 39 (12) : e82
  • [9] Drosophila RecQ5 is required for efficient SSA repair and suppression of LOH in vivo
    Chen, Yixu
    Dui, Wen
    Yu, Zhongsheng
    Li, Changqing
    Ma, Jun
    Jiao, Renjie
    [J]. PROTEIN & CELL, 2010, 1 (05) : 478 - 490
  • [10] DANIELS SB, 1985, GENETICS, V109, P95