BAC transgenic zebrafish for transcriptional promoter and enhancer studies

Methods Mol Biol. 2015;1227:245-58. doi: 10.1007/978-1-4939-1652-8_12.

Abstract

With the advent of BAC recombineering techniques, transcriptional promoter and enhancer isolation studies have become much more feasible in zebrafish than in mouse given the easy access to large numbers of fertilized zebrafish eggs and offspring in general, the easy to follow ex-utero development of zebrafish, an overall less skill demand and a more cost-effective technique. Here we provide guidelines for the generation of BAC recombineering-based transgenic zebrafish for DNA transcriptional promoter and enhancer identification studies as well as protocols for their analysis, which have been successfully applied in our laboratories many times. BAC recombineering in zebrafish allows for economical functional genomics studies, for example by integrating developmental biology with comparative genomics approaches to validate potential enhancer elements of vertebrate transcription factors.

MeSH terms

  • Animals
  • Animals, Genetically Modified*
  • Chromosomes, Artificial, Bacterial / chemistry
  • Chromosomes, Artificial, Bacterial / genetics*
  • Embryo, Nonmammalian
  • Enhancer Elements, Genetic*
  • Female
  • Gene Expression Regulation, Developmental
  • Gene Library
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Lac Operon
  • Male
  • Promoter Regions, Genetic*
  • Transcription, Genetic
  • Zebrafish / genetics*
  • Zebrafish / growth & development
  • Zebrafish / metabolism
  • Zygote

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins