TY - JOUR
T1 - Methylation status of imprinting centers for H19/IGF2 and SNURF/SNRPN in primate embryonic stem cells
AU - Mitalipov, Shoukhrat
AU - Clepper, Lisa
AU - Sritanaudomchai, Hathaitip
AU - Fujimoto, Akihisa
AU - Wolf, Don
PY - 2007/3
Y1 - 2007/3
N2 - Embryonic stem cells (ESCs) hold promise for cell and tissue replacement approaches to treating human diseases based on their capacity to differentiate into a wide variety of somatic cells and tissues. However, long-term in vitro culture and manipulations of ESCs may adversely affect their epigenetic integrity, including imprinting. We have recently reported aberrant biallelic expression of IGF2 and H19 in several rhesus monkey ESC lines, whereas SNRPN and NDN were normally imprinted and expressed predominantly from the paternal allele. The dysregulation of IGF2 and H19 that is associated with tumorigenesis in humans may result from improper maintenance of allele-specific methylation patterns at an imprinting center (IC) upstream of H19. To test this possibility, we performed methylation analysis of several monkey ESC lines by genomic bisulfite sequencing. We investigated methylation profiles of CpG islands within the IGF2/H19 IC harboring the CTCF-6 binding site. In addition, the methylation status of the IC within the promoter/ exon 1 of SNURF/SNRPN known as the Prader-Willi syndrome IC was examined. Our results demonstrate abnormal hypermethylation within the IGF2/H19 IC in all analyzed ESC lines, whereas the SNURF/SNRPN IC was differentially methylated, consistent with monoallelic expression.
AB - Embryonic stem cells (ESCs) hold promise for cell and tissue replacement approaches to treating human diseases based on their capacity to differentiate into a wide variety of somatic cells and tissues. However, long-term in vitro culture and manipulations of ESCs may adversely affect their epigenetic integrity, including imprinting. We have recently reported aberrant biallelic expression of IGF2 and H19 in several rhesus monkey ESC lines, whereas SNRPN and NDN were normally imprinted and expressed predominantly from the paternal allele. The dysregulation of IGF2 and H19 that is associated with tumorigenesis in humans may result from improper maintenance of allele-specific methylation patterns at an imprinting center (IC) upstream of H19. To test this possibility, we performed methylation analysis of several monkey ESC lines by genomic bisulfite sequencing. We investigated methylation profiles of CpG islands within the IGF2/H19 IC harboring the CTCF-6 binding site. In addition, the methylation status of the IC within the promoter/ exon 1 of SNURF/SNRPN known as the Prader-Willi syndrome IC was examined. Our results demonstrate abnormal hypermethylation within the IGF2/H19 IC in all analyzed ESC lines, whereas the SNURF/SNRPN IC was differentially methylated, consistent with monoallelic expression.
KW - Embryonic stem cells
KW - Imprinting
KW - Methylation
KW - Monkey
UR - http://www.scopus.com/inward/record.url?scp=33847621750&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33847621750&partnerID=8YFLogxK
U2 - 10.1634/stemcells.2006-0120
DO - 10.1634/stemcells.2006-0120
M3 - Article
C2 - 17170068
AN - SCOPUS:33847621750
SN - 1066-5099
VL - 25
SP - 581
EP - 588
JO - Stem Cells
JF - Stem Cells
IS - 3
ER -