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Related Experiment Videos

Rapid analysis of gene expression (RAGE) facilitates universal expression profiling.

A Wang1, A Pierce, K Judson-Kremer

  • 1Department of Carcinogenesis, University of Texas M. D. Anderson Cancer Center, PO Box 389, Smithville, TX 78957, USA.

Nucleic Acids Research
|November 11, 1999
PubMed
Summary

A new PCR-based method offers intermediate-scale gene expression analysis, detecting 2-fold changes and identifying new E2F1 targets like Brca1 and Cdk7.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Gene Expression Analysis

Background:

  • Current gene expression analysis methods are limited to single-gene or thousands-of-genes scales.
  • There is a need for flexible, intermediate-scale gene expression profiling.

Purpose of the Study:

  • To develop a rapid, PCR-based method for intermediate-scale gene expression analysis.
  • To enable both directed searches of known genes and surveys of unknown genes.

Main Methods:

  • A novel PCR-based assay adaptable for most eukaryotic genes.
  • Utilizes a single set of reagents and reaction conditions.
  • Suitable for analyzing tens to hundreds of genes across multiple samples.

Main Results:

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  • Reliable detection of gene expression changes (≥2-fold) and high sensitivity (better than 1 in 10,000).
  • Identified new downstream targets of E2F1, including Brca1 and Cdk7, in a transgenic mouse model.
  • Discovered several upregulated unidentified genes and observed changes in apoptosis-related genes.

Conclusions:

  • The developed method provides a flexible and sensitive tool for intermediate-scale gene expression analysis.
  • New insights into E2F1 regulation and potential potentiation of apoptotic pathways in transgenic keratinocytes were gained.