FlEx-Based Transgenic Reporter Lines for Visualization of Cre and Flp Activity in Live Zebrafish

被引:66
作者
Boniface, Emily J. [2 ]
Lu, Jianjun [2 ,3 ]
Victoroff, Tristan [2 ]
Zhu, Meiying [1 ]
Chen, Wenbiao [1 ,2 ]
机构
[1] Vanderbilt Univ, Med Ctr, Dept Mol Physiol & Biophys, Nashville, TN 37232 USA
[2] Oregon Hlth & Sci Univ, Vollum Inst, Portland, OR 97201 USA
[3] Natl Inst Control Pharmaceut & Biol Prod, Natl Ctr Safety Evaluat Drugs, Dept Pathol, Beijing, Peoples R China
关键词
Cre; Flp; CreERT2; reporter line; enhancer trap; fluorescent proteins; tissue-specific Cre; conditional Cre; zebrafish; TOL2 TRANSPOSABLE ELEMENT; REGULATORY ELEMENTS; FLUORESCENT PROTEIN; GENE; EXPRESSION; SYSTEM; RECOMBINATION; MOUSE; TRANSMISSION; SEQUENCE;
D O I
10.1002/dvg.20526
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Site-specific recombinases such as Cre and Flp are invaluable tools for genetic manipulations, but their usage in zebrafish has been limited. Incorporating recently developed flip-excision (FlEx) design that allows stable inversions, we have established zebrafish reporter lines that express bright and ubiquitous EGFP, but switch to express mCherry in the presence of Cre or Flp. Here, we demonstrate the stable inversion in the reporter lines, both in somatic cells and in the germ line by Cre or Flp, and the subsequent reinversion using the other recombinase. Using the reporter lines, we characterized cardiomyocyte-specific Cre lines and neuronal progenitor-specific and tamoxifen-dependent Cre lines. We also used the reporter lines for screening Cre- and Flp-based enhancer trap lines. Similar to the widely used Cre reporter lines in mice, these FlEx-based reporter lines will facilitate the use of recombinases for genetic manipulations in zebrafish. genesis 47:484 491, 2009. (C) 2009 Wiley-Liss, Inc.
引用
收藏
页码:484 / 491
页数:8
相关论文
共 33 条
[21]  
2-M
[22]   Genomewide production of multipurpose alleles for the functional analysis of the mouse genome [J].
Schnütgen, F ;
De-Zolt, S ;
Van Sloun, P ;
Hollatz, M ;
Floss, T ;
Hansen, J ;
Altschmied, J ;
Seisenberger, C ;
Ghyselinck, NB ;
Ruiz, P ;
Chambon, P ;
Wurst, W ;
von Melchner, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (20) :7221-7226
[23]   A directional strategy for monitoring Cre-mediated recombination at the cellular level in the mouse [J].
Schnütgen, F ;
Doerflinger, N ;
Calléja, C ;
Wendling, O ;
Chambon, P ;
Ghyselinck, NB .
NATURE BIOTECHNOLOGY, 2003, 21 (05) :562-565
[24]   Targeting neural circuitry in zebrafish using GAL4 enhancer trapping [J].
Scott, Ethan K. ;
Mason, Lindsay ;
Arrenberg, Aristides B. ;
Ziv, Limor ;
Gosse, Nathan J. ;
Xiao, Tong ;
Chi, Neil C. ;
Asakawa, Kazuhide ;
Kawakami, Koichi ;
Baier, Herwig .
NATURE METHODS, 2007, 4 (04) :323-326
[25]   Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp red fluorescent protein [J].
Shaner, NC ;
Campbell, RE ;
Steinbach, PA ;
Giepmans, BNG ;
Palmer, AE ;
Tsien, RY .
NATURE BIOTECHNOLOGY, 2004, 22 (12) :1567-1572
[26]   Codon-improved Cre recombinase (iCre) expression in the mouse [J].
Shimshek, DR ;
Kim, J ;
Hübner, MR ;
Spergel, DJ ;
Buchholz, F ;
Casanova, E ;
Stewart, AF ;
Seeburg, PH ;
Sprengel, R .
GENESIS, 2002, 32 (01) :19-26
[27]   Generalized lacZ expression with the ROSA26 Cre reporter strain [J].
Soriano, P .
NATURE GENETICS, 1999, 21 (01) :70-71
[28]  
Srinivas S, 2001, BMC Dev Biol, V1, P4, DOI 10.1186/1471-213X-1-4
[29]   Use of FLP/FRT system to study Drosophila development [J].
Theodosiou, NA ;
Xu, T .
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY, 1998, 14 (04) :355-365
[30]   Cre-mediated site-specific recombination in zebrafish embryos [J].
Thummel, R ;
Burket, CT ;
Brewer, JL ;
Sarras, MP ;
Li, L ;
Perry, M ;
McDermott, JP ;
Sauer, B ;
Hyde, DR ;
Godwin, AR .
DEVELOPMENTAL DYNAMICS, 2005, 233 (04) :1366-1377