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Generation of a non‐leaky heat shock–inducible Cre line for conditional Cre/lox strategies in zebrafish
Author(s) -
Hans Stefan,
Freudenreich Dorian,
Geffarth Michaela,
Kaslin Jan,
Machate Anja,
Brand Michael
Publication year - 2011
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.22497
Subject(s) - biology , recombination , zebrafish , microbiology and biotechnology , mcherry , genetics , gene , green fluorescent protein
Cre‐mediated site‐specific recombination has emerged as an indispensable tool for the precise manipulation of the mammalian genome. Recently, we showed that Cre is also highly efficient in zebrafish and temporal control of recombination can be achieved by using the ligand‐inducible CreER T2 . Previous attempts have been made to control recombination by using the temperature inducible hsp70l promoter to conditionally drive the expression of Cre or EGFP‐Cre, respectively. However, in this study we demonstrate that the hsp70l promoter possesses a basal leakiness resulting in Cre‐mediated recombination even at permissive temperatures. In order to prevent non‐conditional recombination, we combined the hsp70l promoter with a mCherry‐tagged ligand‐inducible CreER T2 . At permissive temperatures and in the absence of the ligand tamoxifen (TAM), no non‐conditional recombination is observed indicating tight regulation of CreER T2 . Instead, comprehensive site‐specific recombination is mediated following heat induction and administration of TAM. Developmental Dynamics, 2011. © 2010 Wiley‐Liss, Inc.