ZFIN ID: ZDB-PUB-170818-11
Ligand-Controlled Site-Specific Recombination in Zebrafish
Chekuru, A., Kuscha, V., Hans, S., Brand, M.
Date: 2017
Source: Methods in molecular biology (Clifton, N.J.) 1642: 87-97 (Chapter)
Registered Authors: Brand, Michael, Chekuru, Avinash, Hans, Stefan, Kuscha, Veronika
Keywords: CreERT2, Site-specific recombination, Tamoxifen, Zebrafish
MeSH Terms: none
PubMed: 28815495 Full text @ Methods Mol. Biol.
Cre-mediated site-specific recombination has emerged as an indispensable tool for the precise manipulation of genomes allowing lineage-tracing studies, temporal and spatial misexpressions, and in particular the generation of conditional knockout alleles. Previously, we and others showed that Cre and its ligand-inducible variant CreER(T2) are also highly efficient in the developing and adult zebrafish. The number of Cre driver and effector lines is currently still limited in zebrafish. However, the recent advent of novel genome editing tools such as TALEN and CRISPR/Cas will significantly increase interest in the conditional Cre/lox-technology in this organism. The considerations of basic transgene design and subsequent transgenesis have been addressed elsewhere. Here we outline practical experimental steps for transient functionality tests of CreER(T2) driver and effector constructs. In addition, we introduce detailed protocols to elicit CreER(T2)-mediated recombination in vivo at embryonic as well as adult stages.