ZFIN ID: ZDB-PUB-151119-16
Generation of a conditional lima1a allele in zebrafish using the FLEx switch technology
Jungke, P., Hans, S., Gupta, M., Machate, A., Zöller, D., Brand, M.
Date: 2016
Source: Genesis (New York, N.Y. : 2000) 54(1): 19-28 (Journal)
Registered Authors: Brand, Michael, Gupta, Mansi, Hans, Stefan, Jungke, Peggy, Machate, Anja
Keywords: FLEx switch, Lima1a, conditional alleles, zebrafish
MeSH Terms: Alleles*; Animals; Animals, Genetically Modified; Cytoskeletal Proteins/genetics; DNA Nucleotidyltransferases/chemistry (all 13) expand
PubMed: 26572123 Full text @ Genesis
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ABSTRACT
Gene trapping has emerged as a valuable tool to create conditional alleles in various model organisms. Here we report the FLEx-based gene trap vector SAGFLEx that allows the generation of conditional mutations in zebrafish by gene-trap mutagenesis. The SAGFLEx gene trap cassette comprises the rabbit β-globin splice acceptor and the coding sequence of GFP, flanked by pairs of inversely oriented heterotypic target sites for the site-specific recombinases Cre and Flp. Insertion of the gene-trap cassette into endogenous genes can result in conditional mutations that are stably inverted by Cre and Flp, respectively. To test the functionality of this system we performed a pilot screen and analyzed the insertion of the gene-trap cassette into the lima1a gene locus. In this lima1a allele, GFP expression faithfully recapitulated the endogenous lima1a expression and resulted in a complete knockout of the gene in homozygosity. Application of either Cre or Flp was able to mediate the stable inversion of the gene trap cassette and showed the ability to conditionally rescue or reintroduce the gene inactivation. Combined with pharmacologically inducible site specific recombinases the SAGFLEx vector insertions will enable precise conditional knockout studies in a spatial- and temporal-controlled manner. This article is protected by copyright. All rights reserved.
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