Korean J Reprod Med Search

CLOSE


Korean Journal of Reproductive Medicine 2010;37(1):25-31.
Published online March 1, 2010.
Epigenetic Study of XIST Gene from Female and Male Cells by Pyrosequencing.
Hwan Hee Kim, Yeo Jin Yun, Min Ae Song, Suman Lee
Functional Genomics Lab, CHA Stem Cell Research Institute, School of Medicine, CHA University, Bundang-Gu, Sungnam-Si, Kyunggi-Do, 463-836, Korea.
Abstract
OBJECTIVE
X inactivation is the silencing one of the two X chromosomes in female mammals for gene dosage on the X-chromosome between female and male. X inactivation is controlled by X inactive-specific transcript (XIST) gene, untranslated RNA. XIST is expressed only from the inactive X (Xi), not expressed from the active X (Xa). The Xist promoter is methylated on the silent Xist allele on the Xa in somatic cells, and less methylated on the Xist-expressing Xi. We investigated the difference of XIST methylation pattern of the promoter and 5'-region of XIST from male (XY) and female (XX) subjects. METHODS: The direct quantification of XIST methylation is required for clinical application of normal XX and XY blood. Methylation percentage of eight CpG sites (-1696, -1679, -1475, -1473, -1469, +947, +956, +971) of XIST gene were diagnosed by pyrosequencing. RESULTS: We directly quantitated the methylation percentage of the promoter and 5'-end of XIST by pyrosequencing. The average methylation percentages at CpG6-8 sites (+947, +956, +971) were 45.2% at CpG6, 49.9% at CpG7, and 44.2% at CpG8 from normal female and normal male were 90.6%, 96.7%, 87.8%, respectively. Nether CpG 1-5sites (-1696, -1679, -1475, -1473, -1469) had any effect on XX and XY. CONCLUSION: This method is sensitive for quantifying the small percentage change in the methylation status of XIST, and may be used for diagnosis.
Key Words: XIST; CpG; DNA methylation; Pyrosequencing
TOOLS
Share :
Facebook Twitter Linked In Google+ Line it
METRICS Graph View
  • 541 View
  • 2 Download
Related articles in Clin Exp Reprod Med


ABOUT
ARTICLE CATEGORY

Browse all articles >

BROWSE ARTICLES
AUTHOR INFORMATION
Editorial Office
Department of Obstetrics and Gynecology CHA Bundang Medical Center, CHA University
59 Yatap-ro, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea
Tel: +82-31-727-8701    CP: +82-10-9072-3154    E-mail: hwas0605@cha.ac.kr                

Copyright © 2022 by Korean Society for Reproductive Medicine.

Developed in M2PI

Close layer
prev next