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An Ecdysone Receptor-based Singular Gene Switch for Deliberate Expression of Transgene with Robustness, Reversibility, and Negligible Leakiness
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Transient Transgenics: An Efficient Method to Identify Gene Regulatory Elements.

Warren E Zimmer1, Robert J Schwartz2

  • 1Department of Medical Physiology, Texas A&M College of Medicine, Bryan, TX, USA. wezimmer@tamu.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a new method using Bacterial Artificial Chromosome (BAC) DNA transfer in mouse embryos to find DNA sequences controlling cell-specific gene expression during development.

Keywords:
Bacterial artificial chromosomes (BAC)Red E/T recombineeringTransient transgenicVISTAΒ-galactosidase

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Area of Science:

  • Developmental Biology
  • Molecular Genetics
  • Genomics

Background:

  • Identifying cis-acting DNA sequences is crucial for understanding gene regulation.
  • Cell-specific gene expression patterns are essential for embryonic development.

Purpose of the Study:

  • To develop an efficient method for identifying cis-acting DNA sequences.
  • To pinpoint regulatory elements driving cell-specific gene expression in developing embryos.

Main Methods:

  • Transfer of modified Bacterial Artificial Chromosome (BAC) genomic DNAs with a reporter gene into mouse embryos.
  • In utero development of injected embryos followed by collection and analysis.
  • Utilizing the LacZ reporter gene for microscopy and sectioning to analyze gene activity.

Main Results:

  • The method successfully identifies cis-element activity in developing mouse embryos.
  • Analysis of reporter gene expression reveals cell-specific patterns.
  • Facilitates evaluation of cis-element function through subsequent mutational analysis.

Conclusions:

  • This novel technique provides an efficient way to identify gene regulatory domains.
  • It enables the discovery of sequences specifying stage-specific gene expression.
  • The method is valuable for studying developmental gene regulation.