Identification, cloning and expression analysis of the pluripotency promoting Nanog genes in mouse and human

Abstract

The murine Nanog gene, a member of the homeobox family of DNA binding transcription factors, has been shown recently to maintain pluripotency of embryonic stem cells. We have used a sequence homology and expression screen to identify and clone the mouse and human Nanog genes and characterized their phylogenetic context and expression patterns. We report here the gene structure and expression patterns of the mouse Nanog gene, the human Nanog and Nanog2 genes, and six processed human Nanog pseudogenes. Mouse Nanog expression is high in undifferentiated embryonic stem cells and is down-regulated during embryonic stem cell differentiation, concomitant with loss of pluripotency. Murine embryonic Nanog expression is detected in the inner cell mass of the blastocyst. After implantation, Nanog is detectable at embryonic day (E) 6 in proximal epiblast in the region of the presumptive primitive streak. Expression extends distally as the streak elongates during gastrulation and remains restricted to epiblast. Nanog RNA is down-regulated in cells ingressing through the streak to form mesoderm and definitive endoderm. Nanog expression also marks the pluripotent germ cells of the nascent gonad at E11.5-E12.5 and is highly expressed in germ cell tumour and teratoma-derived cell lines. Reverse transcriptase-polymerase chain reaction analysis detected mouse Nanog expression at low levels in several adult tissues. The human Nanog genes are expressed in embryonic stem cells and down-regulated in all adult tissues and differentiated cell lines examined. High levels of human Nanog expression were detected by Northern analysis in the undifferentiated N-Tera embryonal carcinoma cell line. The conservation in gene sequence, structure, and expression of mouse and human Nanog and Nanog2 genes may reflect a common role in the maintenance of pluripotency in both species. Developmental

Research of cardiomyocyte precursors in adult rat heart

“…The different expression of CD9, nanog and taube nuss that we observed in adult myocardium could be associated to the degree of differentiation of embryonic stem cells. Indeed, it has been reported that nanog and CD9 expression was elevated in undifferentiated embryonic stem cells and down-regulated during their differentiation, in concomitance with the loss of the pluripotency (Cui et al, 2004;Hart et al, 2004). Since it has been already demonstrated that the absence of taube nuss causes apoptosis of stem cells from blastocisty inner cellular mass (Voss et al, 2000), we postulate that this factor could be important for stem cell survival even in adult heart.…”

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

“…The period of loss of NANOG expression occurring at peri‐implantation (Chambers et al , 2003; Acampora et al , 2013) may enable cells of the epiblast to downregulate a number of pivotal naïve pluripotency determinants, including ESRRB (Adachi et al , 2013). In the post‐implantation epiblast, NANOG is re‐expressed (Hart et al , 2004; Osorno et al , 2012; Hoffman et al , 2013) but ESRRB is not (Adachi et al , 2013), likely due to differences in signaling environments between pre‐ and post‐implantation epiblasts. It is noteworthy that subpopulations of ESC cultures have been proposed to bear a similar character to primed pluripotent cells and vice versa (Hayashi et al , 2008; Han et al , 2010).…”

Assessing Self-Renewal and Differentiation in Human Embryonic Stem Cell Lines

“…Oct4 and Nanog are essential for the maintenance of pluripotency in both hESCs and mESCs, and knockout or knockdown of either gene causes differentiation [38,39]. Oct4 and Nanog are, however, not uniquely expressed in undifferentiated hESCs: Oct4 is expressed in germ cells, and Nanog has recently been reported to be expressed in mature tissues [40,41]. Moreover, the expression of Oct4 declines slowly as cells differentiate, and the change in levels is small.…”