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Related Concept Videos

Complementation Tests00:49

Complementation Tests

A complementation test is a simple cross to identify whether the two mutations are located on the same gene or different genes. It was first performed by Edward Lewis in the 1940s while working on fruit flies. He developed the test to identify the location and arrangement of different mutations on chromosomes.
Organisms heterozygous for different mutations are crossed pairwise in all combinations. If present on different genes, the mutations can complement each other by providing the missing...

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Systematic identification of cis-silenced genes by trans complementation.

Jae Hyun Lee1, Branimir Bugarija, Enrique J Millan

  • 1Department of Human Genetics, Howard Hughes Medical Institute, University of Chicago, Chicago, IL 60637, USA.

Human Molecular Genetics
|December 4, 2008
PubMed
Summary

Trans complementation reveals an

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Gene transcription is regulated by cis-acting chromatin states and trans-acting cellular factors.
  • Dissecting the relative contributions of these regulatory mechanisms is crucial for understanding cell fate.
  • Existing methods struggle to isolate cis from trans regulatory effects.

Purpose of the Study:

  • To develop a novel assay, termed trans complementation, for dissecting cis- versus trans-regulatory mechanisms in gene expression.
  • To identify genes regulated by cis-acting mechanisms that are unresponsive to the cellular environment.
  • To investigate the role of such unresponsive gene states in cell fate determination.

Main Methods:

  • Fusing two distinct cell types to create a shared cellular environment (trans complementation).
  • Analyzing differential gene expression between the two genomes within the fused cell.
  • Identifying genes exhibiting altered expression patterns attributable to cis-acting factors.

Main Results:

  • A novel state of transcriptional competency, termed 'occluded,' was identified.
  • Occluded genes are silenced by cis-acting mechanisms, rendering them unresponsive to trans-acting factors.
  • These occluded genes often control alternative cell fates and are stably maintained during cell division.
  • The assay provides a functional readout of chromatin state distinct from gene expression.

Conclusions:

  • Gene occlusion by cis-acting mechanisms is a key factor in restricting cell fate.
  • Trans complementation is a powerful tool for distinguishing cis and trans regulatory effects.
  • This work offers new insights into the stability and function of chromatin-mediated gene silencing.