title: Detection of allele-specific transcripts by the polymerase chain reaction (AST-PCR).
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.
authors.affiliation: MGC-Department of Human Genetics, Leiden University, The Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Top
authors.forename: N
authors.initials: N
authors.lastname: van den Boorn
authors.forename: A
authors.initials: A
authors.lastname: van der Zee
authors.forename: L M
authors.initials: LM
authors.lastname: Havekes
authors.forename: R R
authors.initials: RR
authors.lastname: Frants
mesh_terms: D000483:Alleles; D001483:Base Sequence; D004247:DNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D006580:Genetic Carrier Screening; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000235:genetics*; D007962:Leukocytes / Q000502:physiology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012313:RNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D011973:Receptors, LDL / Q000235:genetics*; D012016:Reference Values; D015183:Restriction Mapping; D014158:Transcription, Genetic
chemical_list: D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011973:Receptors, LDL; D012313:RNA; D004247:DNA