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a Centre for Stem Cell Biology and Developmental Genetics and
b Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne, United Kingdom;
c School of Biological and Biomedical Sciences, University of Durham, Durham, United Kingdom;
d Newcastle Fertility Centre at Life, International Centre for Life, Newcastle upon Tyne, United Kingdom
Key Words. NANOG • Human embryonic stem cells • Pluripotency • Small interfering RNA • Trophectoderm • Primitive endoderm
Correspondence: Majlinda Lako, Ph.D., Centre for Stem Cell Biology and Developmental Genetics, University of Newcastle International Centre for Life, Newcastle NE1 3BZ, United Kingdom. Telephone: 0191-241-8688; Fax: 0191-241-8666; e-mail: Majlinda.Lako{at}ncl.ac.uk
The homeobox transcription factor Nanog has been proposed to play a crucial role in the maintenance of the undifferentiated state of murine embryonic stem cells. A human counterpart, NANOG, has been identified, but its function and localization have not hitherto been described. We have used a combination of RNA interference and quantitative real-time polymerase chain reaction to study NANOG in human embryonic stem and embryonic carcinoma cells. Transfection of NANOG-specific small interfering RNAs reduced levels of NANOG transcript and protein and induced activation of the extraembryonic endoderm-associated genes GATA4, GATA6, LAMININ B1, and AFP as well as upregulation of trophectoderm-associated genes CDX2, GATA2, hCG-alpha, and hCG-beta. Immunostaining of preimplantation human embryos showed that NANOG was expressed in the inner cell mass of expanded blastocysts but not in earlier-stage embryos, consistent with a role in the maintenance of pluripotency. Taken together, our findings suggest that NANOG acts as a gatekeeper of pluripotency in human embryonic stem and carcinoma cells by preventing their differentiation to extraembryonic endoderm and trophectoderm lineages.
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