Temporally-Controlled Site-Specific Recombination in Zebrafish

被引:138
作者
Hans, Stefan [1 ,2 ]
Kaslin, Jan [1 ,2 ]
Freudenreich, Dorian [1 ,2 ]
Brand, Michael [1 ,2 ]
机构
[1] Tech Univ Dresden, Ctr Biotechnol, D-8027 Dresden, Germany
[2] Tech Univ Dresden, Ctr Regenerative Therapies Dresden, D-8027 Dresden, Germany
关键词
D O I
10.1371/journal.pone.0004640
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Conventional use of the site-specific recombinase Cre is a powerful technology in mouse, but almost absent in other vertebrate model organisms. In zebrafish, Cre-mediated recombination efficiency was previously very low. Here we show that using transposon-mediated transgenesis, Cre is in fact highly efficient in this organism. Furthermore, temporal control of recombination can be achieved by using the ligand-inducible CreER(T2). Site-specific recombination only occurs upon administration of the drug tamoxifen (TAM) or its active metabolite, 4-hydroxy-tamoxifen (4-OHT). Cre-mediated recombination is detectable already 4 or 2 hours after administration of TAM or 4-OHT, demonstrating fast recombination kinetics. In addition, low doses of TAM allow mosaic labeling of single cells. Combined, our results show that conditional Cre/lox will be a valuable tool for both, embryonic and adult zebrafish studies. Furthermore, single copy insertion transgenesis of Cre/lox constructs suggest a strategy suitable also for other organisms.
引用
收藏
页数:7
相关论文
共 30 条
[1]   Genetic dissection of neural circuits by Tol2 transposon-mediated Gal4 gene and enhancer trapping in zebrafish [J].
Asakawa, Kazuhide ;
Suster, Maximiliano L. ;
Mizusawa, Kanta ;
Nagayoshi, Saori ;
Kotani, Tomoya ;
Urasaki, Akihiro ;
Kishimoto, Yasuyuki ;
Hibi, Masahiko ;
Kawakami, Koichi .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (04) :1255-1260
[2]   Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases [J].
Doyon, Yannick ;
McCammon, Jasmine M. ;
Miller, Jeffrey C. ;
Faraji, Farhoud ;
Ngo, Catherine ;
Katibah, George E. ;
Amora, Rainier ;
Hocking, Toby D. ;
Zhang, Lei ;
Rebar, Edward J. ;
Gregory, Philip D. ;
Urnov, Fyodor D. ;
Amacher, Sharon L. .
NATURE BIOTECHNOLOGY, 2008, 26 (06) :702-708
[3]  
Driever W, 1996, DEVELOPMENT, V123, P37
[4]   Molecular neuroanatomy's "three Gs": A primer [J].
Dymecki, Susan M. ;
Kim, Jun Chul .
NEURON, 2007, 54 (01) :17-34
[5]   Mifepristone-inducible LexPR system to drive and control gene expression in transgenic zebrafish [J].
Emelyanov, Alexander ;
Parinov, Serguei .
DEVELOPMENTAL BIOLOGY, 2008, 320 (01) :113-121
[6]  
Feil R, 2007, Handb Exp Pharmacol, P3
[7]   Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding domains [J].
Feil, R ;
Wagner, J ;
Metzger, D ;
Chambon, P .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 237 (03) :752-757
[8]   Heat-shock induction of T-cell lymphoma/leukaemia in conditional Cre/lox-regulated transgenic zebrafish [J].
Feng, Hui ;
Langenau, David M. ;
Madge, Jennifer A. ;
Quinkertz, Andre ;
Gutierrez, Alejandro ;
Neuberg, Donna S. ;
Kanki, John P. ;
Look, A. Thomas .
BRITISH JOURNAL OF HAEMATOLOGY, 2007, 138 (02) :169-175
[9]  
Haffter P, 1996, DEVELOPMENT, V123, P1
[10]   Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: A tool for temporally regulated gene activation/inactivation in the mouse [J].
Hayashi, S ;
McMahon, AP .
DEVELOPMENTAL BIOLOGY, 2002, 244 (02) :305-318