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Related Experiment Videos

RNA-FISH to analyze allele-specific expression.

G Braidotti1

  • 1Department of Cell Biology and Genetics, Erasmus University, Rotterdam, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)
|July 5, 2003
PubMed
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Analyzing imprinted gene expression requires distinguishing maternal and paternal gene copies. The human IGF2R gene shows polymorphic monoallelic expression, suggesting it may not be consistently imprinted across all individuals.

Area of Science:

  • Genetics
  • Molecular Biology
  • Epigenetics

Background:

  • Distinguishing RNA synthesis from maternal versus paternal alleles is crucial for analyzing imprinted gene expression.
  • Allele-specific expression studies often rely on polymorphisms and bulk cell RNA analysis.

Purpose of the Study:

  • To investigate the allele-specific expression of the human IGF2R gene.
  • To determine if the human IGF2R gene exhibits imprinted expression patterns.

Main Methods:

  • Utilized restriction fragment length polymorphism (RFLP) analysis.
  • Employed reverse transcriptase polymerase chain reaction (RT-PCR) in heterozygotes.
  • Analyzed RNA from lymphoblastoid cell samples.

Main Results:

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  • The human IGF2R gene has a hemimethylated intronic CpG island, similar to the mouse imprinting box.
  • IGF2R mRNA was detected from both maternal and paternal alleles in most samples.
  • One sample showed paternal allele reactivation during cell culturing.

Conclusions:

  • Monoallelic expression of the human IGF2R gene appears to be a polymorphic trait.
  • A small minority of individuals may exhibit monoallelic expression.
  • The imprinting status of the human IGF2R gene remains a question.