Localization of trinucleotide repeat sequences in myotonic dystrophy cells using a single fluorochrome-labeled PNA probe.
A labeled peptide nucleic acid (PNA) antisense probe was used to study the spatial distribution of triplet repeats (CTG) in human myotonic dystrophy (DM) cells by high-resolution fluorescence in situ hybridization (FISH). It was found that transcripts containing triplet repeats were present as a number of discrete foci in the DM nuclei. Greater numbers of foci were visible with the PNA probe than a comparable DNA probe. The PNA probe was also used to visualize the triplet-repeat expansion within the DM gene located on chromosome 19. Using the intensity of the expanded triplet-repeat on the chromosomes as a reference, it was estimated there were between 15-230 RNA molecules in each focus observed in DM nuclei.
Taneja
K L
KL
University of Massachusetts Medical Center, Worcester, USA.
D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics / Q000473:pathology; D002877:Chromosomes, Human / Q000737:chemistry; D018628:Gene Dosage; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D009223:Myotonic Dystrophy / Q000235:genetics* / Q000473:pathology; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D015345:Oligonucleotide Probes / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D010455:Peptides / Q000235:genetics; D018911:Trinucleotide Repeats / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D016376:Oligonucleotides, Antisense; D010455:Peptides