The use of in ovo electroporation for the rapid analysis of neural-specific murine enhancers

John Timmer, Jane Johnson, Lee Niswander

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

The identification and characterization of DNA sequences necessary for proper gene expression have provided insights into gene regulation and generated tools useful for further experimentation. Studies of developmentally regulated genes have demonstrated how transcription factors interact at enhancers to generate restricted patterns of expression during embryogenesis. In vertebrates, the pursuit of such studies has relied on the generation of transgenic mice and thus has been limited by the time and expense required generating and characterizing these mice. The recently developed technique of in ovo electroporation allows the rapid introduction of exogenous DNA into developing chicken embryos. Here we have used this technique to introduce DNA containing murine enhancer/reporter constructs into cells of the chicken neural tube, resulting in appropriate expression of the reporter. This technique has the potential to greatly reduce the effort involved in the study of vertebrate enhancers. Furthermore, we have characterized factors such as timing of electroporation, concentration of DNA, and choice of basal promoters and found that they can influence the degree to which expression of enhancer constructs reflects endogenous gene expression.

Original languageEnglish (US)
Pages (from-to)123-132
Number of pages10
JournalGenesis
Volume29
Issue number3
DOIs
StatePublished - 2001

Fingerprint

Electroporation
Vertebrates
Chickens
DNA
Gene Expression
Neural Tube
Transgenic Mice
Genes
Embryonic Development
Transcription Factors
Embryonic Structures

Keywords

  • bHLH
  • Chicken
  • Electroporation
  • Enhancer analysis
  • Neural

ASJC Scopus subject areas

  • Genetics

Cite this

The use of in ovo electroporation for the rapid analysis of neural-specific murine enhancers. / Timmer, John; Johnson, Jane; Niswander, Lee.

In: Genesis, Vol. 29, No. 3, 2001, p. 123-132.

Research output: Contribution to journalArticle

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