Detection of allele-specific transcripts by the polymerase chain reaction (AST-PCR)

Biochem Biophys Res Commun. 1991 Aug 15;178(3):1319-25. doi: 10.1016/0006-291x(91)91038-e.

Abstract

We have applied the polymerase chain reaction to detect differences in relative amount of allele-specific transcripts of the low density lipoprotein receptor gene in individuals with heterozygous familial hypercholesterolemia (FH). This method is based on detection of loss of heterozygosity of exon-specific restriction fragment length polymorphisms present in amplified mRNA fragments as compared to amplified genomic DNA fragments. We detected loss of allele-specific mRNA in 20% of informative FH patients. In principle, this method enables the analysis of relative allele-specific transcript levels of any expressed gene and can thus be generally applied to study processes involving differential gene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles*
  • Base Sequence
  • DNA / blood
  • DNA / genetics
  • DNA / isolation & purification
  • Genetic Carrier Screening
  • Humans
  • Hyperlipoproteinemia Type II / genetics*
  • Leukocytes / physiology
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length*
  • RNA / blood
  • RNA / genetics
  • RNA / isolation & purification
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics*
  • Receptors, LDL / genetics*
  • Reference Values
  • Restriction Mapping
  • Transcription, Genetic*

Substances

  • Oligonucleotide Probes
  • RNA, Messenger
  • Receptors, LDL
  • RNA
  • DNA