Conditional gene-trap mutagenesis in zebrafish.

Maddison LA, Li M, Chen W
Methods Mol Biol. 2014 1101: 393-411

PMID: 24233792 · PMCID: PMC4129463 · DOI:10.1007/978-1-62703-721-1_19

Zebrafish has become a widely used model for analysis of gene function. Several methods have been used to create mutations in this organism and thousands of mutant lines are available. However, all the conventional zebrafish mutations affect the gene in all cells at all time, making it difficult to determine tissue-specific functions. We have adopted a FlEx Trap approach to generate conditional mutations in zebrafish by gene-trap mutagenesis. Combined with appropriate Cre or Flp lines, the insertional mutants not only allow spatial- and temporal-specific gene inactivation but also permit spatial- and temporal-specific rescue of the disrupted gene. We provide experimental details on how to generate and use such mutations.

MeSH Terms (13)

Alleles Animals Base Sequence DNA Transposable Elements Embryo Culture Techniques Female Genetic Vectors Male Microinjections Mutagenesis, Insertional Organ Specificity Phenotype Zebrafish

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