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Testing transgenic regulatory elements through live mouse imaging.

Naama Klopstock1, Carol Levy, Devorah Olam

  • 1Goldyne Savad Institute of Gene Therapy, Hadassah - Hebrew University Medical Center, P.O.B. 12000, Jerusalem 91120, Israel. Naamitk@gmail.com

FEBS Letters
|August 4, 2007
PubMed
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Researchers developed a universal cloning cassette for in vivo assessment of regulatory elements. This method allows rapid screening of transgenic mice, revealing that some regulatory elements like the Island Element were not beneficial for transgene expression.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Transgene expression is often affected by genomic position and DNA methylation.
  • Efficient methods are needed to assess regulatory elements in vivo.
  • Luciferase reporter systems are valuable tools for monitoring gene expression.

Purpose of the Study:

  • To develop a universal cloning cassette for in vivo assessment of regulatory elements.
  • To evaluate the efficacy of specific regulatory elements, including insulators and demethylation elements, in controlling transgene expression.
  • To establish a rapid and cost-effective screening method for transgenic founders.

Main Methods:

  • Development of a universal cloning cassette containing a luciferase reporter gene.
  • Utilizing a cooled, coupled-device camera (CCCD) for in vivo imaging of luciferase expression in live mice.

Related Experiment Videos

  • Generating and analyzing transgenic mice carrying engineered regulatory elements.

    • Main Results:

    • The developed system enabled quick and inexpensive screening of transgenic founders.
    • The chicken beta-globin 5'HS4 insulator did not consistently ensure the desired expression pattern.
    • The Island Element, intended for demethylation, did not provide a beneficial effect on transgene expression.
    • Both liver-specific and developmentally regulated promoters generally showed expected expression patterns.

    Conclusions:

    • The universal cloning cassette is effective for in vivo assessment of regulatory elements in transgenic mice.
    • The study highlights the variable performance of specific regulatory elements in controlling transgene expression.
    • This approach facilitates efficient screening and characterization of regulatory elements for gene expression studies.