Prader-Willi and Angelman syndromes

Diagnosis with a bisulfite-treated methylation-specific PCR method

Kenjiro Kosaki, M. J. McGinniss, A. N. Veraksa, W. J. McGinnis, K. L. Jones

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

The putative promoter region of the SNRPN gene contains a CpG island which is heavily methylated in the maternally derived allele and unmethylated in the paternally derived allele. In patients with Prader-Willi syndrome (PWS) only the methylated allele is present, while in those with Angelman syndrome (AS) only the unmethylated allele is present. The purpose of this paper is to report a polymerase chain reaction (PCR)-based assay to evaluate methylation status of the CpG island of the SNRPN gene and to show that this assay allows rapid diagnosis of PWS and AS. Methylated cytosines in the CpG dinucleotide are resistant to chemical modification by sodium bisulfite. In contrast, bisulfite treatment converts all unmethylated cytosines to uracil. Based on this differential effect, the bisulfite-modified DNA sequence of a methylated allele was successfully distinguished from that of an unmethylated allele using 2 sets of allele-specific primer pairs: a methylated allele- specific primer pair (MET) and an unmethylated allele-specific primer pair (UN-MET). Bisulfite-modified DNA from 10 patients with PWS amplified only with the MET pair while modified DNA from 5 patients with AS amplified only with the UN-MET pair. Modified DNA from 50 normal unrelated individuals amplified with both primer pairs. In that methylation-specific PCR (MSPCR) can detect all presently test-able causes of PWS and AS in a rapid and cost- effective fashion, serious consideration should be given to the use of this test in the initial evaluation of all patients in which PWS or AS is being considered.

Original languageEnglish
Pages (from-to)308-313
Number of pages6
JournalAmerican Journal of Medical Genetics
Volume73
Issue number3
DOIs
Publication statusPublished - 1997
Externally publishedYes

Fingerprint

Angelman Syndrome
Prader-Willi Syndrome
Methylation
Alleles
Polymerase Chain Reaction
snRNP Core Proteins
CpG Islands
United Nations
Cytosine
DNA
hydrogen sulfite
Uracil
Genetic Promoter Regions
Genes

Keywords

  • Angelman syndrome
  • CpG island
  • Methylation
  • Prader-Willi syndrome
  • SNRPN

ASJC Scopus subject areas

  • Genetics(clinical)

Cite this

Prader-Willi and Angelman syndromes : Diagnosis with a bisulfite-treated methylation-specific PCR method. / Kosaki, Kenjiro; McGinniss, M. J.; Veraksa, A. N.; McGinnis, W. J.; Jones, K. L.

In: American Journal of Medical Genetics, Vol. 73, No. 3, 1997, p. 308-313.

Research output: Contribution to journalArticle

Kosaki, Kenjiro ; McGinniss, M. J. ; Veraksa, A. N. ; McGinnis, W. J. ; Jones, K. L. / Prader-Willi and Angelman syndromes : Diagnosis with a bisulfite-treated methylation-specific PCR method. In: American Journal of Medical Genetics. 1997 ; Vol. 73, No. 3. pp. 308-313.
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