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SOX17 is a critical specifier of human primordial germ cell fate.

Irie N, Weinberger L, Tang WW, Kobayashi T, Viukov S, Manor YS, Dietmann S, Hanna JH, Surani MA - Cell (2014)

Bottom Line: Here, we demonstrate specification of hPGC-like cells (hPGCLCs) from germline competent pluripotent stem cells.Remarkably, SOX17 is the key regulator of hPGC-like fate, whereas BLIMP1 represses endodermal and other somatic genes during specification of hPGCLCs.We have established a foundation for future studies on resetting of the epigenome in hPGCLCs and hPGCs for totipotency and the transmission of genetic and epigenetic information.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Cancer Research UK Gurdon Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 1QN, UK; Department of Physiology, Development and Neuroscience, Downing Street, University of Cambridge, Cambridge CB2 3EG, UK; Wellcome Trust-Medical Research Council Stem Cell Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 3EG, UK.

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Generation of SOX17 KO in WIS2-NANOS3-mCherry hESC Line, Related to Figure 6(A) Expression analysis by RT-qPCR of pluripotency and germ cell genes after knock-down of SOX17 in TCam-2. Two independent, miRs against SOX17 (S17 miR #1 and #2) were used for the knockdown with a scramble miR as control. Error bars are mean ± SD. Relative expression levels are shown with normalization to GAPDH. Representative data were shown from two independent biological replicates.(B) Targeting strategy of SOX17 knockout in hESC with the designated guide RNA (gRNA) and the resulting deleted sequences.(C) Immunofluorescence of SOX17, BLIMP1 and TFAP2C (left panel); and TFAP2C and NANOS3-mCherry on wild-type (WT) and SOX17 KO day 4 embryoids. Scale bars = 70 μm(D) FACS analysis of day 4 embryoids derived from wild-type (WT), SOX17 knockout (SOX17 KO) and 1 to 1 mixture of WT and SOX17 KO cells (WT + SOX17 KO).
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figs6: Generation of SOX17 KO in WIS2-NANOS3-mCherry hESC Line, Related to Figure 6(A) Expression analysis by RT-qPCR of pluripotency and germ cell genes after knock-down of SOX17 in TCam-2. Two independent, miRs against SOX17 (S17 miR #1 and #2) were used for the knockdown with a scramble miR as control. Error bars are mean ± SD. Relative expression levels are shown with normalization to GAPDH. Representative data were shown from two independent biological replicates.(B) Targeting strategy of SOX17 knockout in hESC with the designated guide RNA (gRNA) and the resulting deleted sequences.(C) Immunofluorescence of SOX17, BLIMP1 and TFAP2C (left panel); and TFAP2C and NANOS3-mCherry on wild-type (WT) and SOX17 KO day 4 embryoids. Scale bars = 70 μm(D) FACS analysis of day 4 embryoids derived from wild-type (WT), SOX17 knockout (SOX17 KO) and 1 to 1 mixture of WT and SOX17 KO cells (WT + SOX17 KO).

Mentions: Expression of SOX17 among T-positive cells prior to BLIMP1 apparently marks the onset of hPGCLC specification, which is a key difference between the specification of human and mouse germline fate (see Figure 4). Notably, SOX17 and BLIMP1 are also expressed in the authentic in vivo hPGCs and in TCam-2 (de Jong et al., 2008) (Figure 2). Knockdown of SOX17 in TCam-2, which exhibits key germ cell characteristics (Looijenga et al., 2014) (Figure 2), induced repression of the pluripotency genes NANOG, as well as of the PGC-genes BLIMP1, NANOS3, TFAP2C, STELLA, and KIT (Figure S6A). This suggests that SOX17 might be important for regulating the established germline gene expression network.


SOX17 is a critical specifier of human primordial germ cell fate.

Irie N, Weinberger L, Tang WW, Kobayashi T, Viukov S, Manor YS, Dietmann S, Hanna JH, Surani MA - Cell (2014)

Generation of SOX17 KO in WIS2-NANOS3-mCherry hESC Line, Related to Figure 6(A) Expression analysis by RT-qPCR of pluripotency and germ cell genes after knock-down of SOX17 in TCam-2. Two independent, miRs against SOX17 (S17 miR #1 and #2) were used for the knockdown with a scramble miR as control. Error bars are mean ± SD. Relative expression levels are shown with normalization to GAPDH. Representative data were shown from two independent biological replicates.(B) Targeting strategy of SOX17 knockout in hESC with the designated guide RNA (gRNA) and the resulting deleted sequences.(C) Immunofluorescence of SOX17, BLIMP1 and TFAP2C (left panel); and TFAP2C and NANOS3-mCherry on wild-type (WT) and SOX17 KO day 4 embryoids. Scale bars = 70 μm(D) FACS analysis of day 4 embryoids derived from wild-type (WT), SOX17 knockout (SOX17 KO) and 1 to 1 mixture of WT and SOX17 KO cells (WT + SOX17 KO).
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Related In: Results  -  Collection

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figs6: Generation of SOX17 KO in WIS2-NANOS3-mCherry hESC Line, Related to Figure 6(A) Expression analysis by RT-qPCR of pluripotency and germ cell genes after knock-down of SOX17 in TCam-2. Two independent, miRs against SOX17 (S17 miR #1 and #2) were used for the knockdown with a scramble miR as control. Error bars are mean ± SD. Relative expression levels are shown with normalization to GAPDH. Representative data were shown from two independent biological replicates.(B) Targeting strategy of SOX17 knockout in hESC with the designated guide RNA (gRNA) and the resulting deleted sequences.(C) Immunofluorescence of SOX17, BLIMP1 and TFAP2C (left panel); and TFAP2C and NANOS3-mCherry on wild-type (WT) and SOX17 KO day 4 embryoids. Scale bars = 70 μm(D) FACS analysis of day 4 embryoids derived from wild-type (WT), SOX17 knockout (SOX17 KO) and 1 to 1 mixture of WT and SOX17 KO cells (WT + SOX17 KO).
Mentions: Expression of SOX17 among T-positive cells prior to BLIMP1 apparently marks the onset of hPGCLC specification, which is a key difference between the specification of human and mouse germline fate (see Figure 4). Notably, SOX17 and BLIMP1 are also expressed in the authentic in vivo hPGCs and in TCam-2 (de Jong et al., 2008) (Figure 2). Knockdown of SOX17 in TCam-2, which exhibits key germ cell characteristics (Looijenga et al., 2014) (Figure 2), induced repression of the pluripotency genes NANOG, as well as of the PGC-genes BLIMP1, NANOS3, TFAP2C, STELLA, and KIT (Figure S6A). This suggests that SOX17 might be important for regulating the established germline gene expression network.

Bottom Line: Here, we demonstrate specification of hPGC-like cells (hPGCLCs) from germline competent pluripotent stem cells.Remarkably, SOX17 is the key regulator of hPGC-like fate, whereas BLIMP1 represses endodermal and other somatic genes during specification of hPGCLCs.We have established a foundation for future studies on resetting of the epigenome in hPGCLCs and hPGCs for totipotency and the transmission of genetic and epigenetic information.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Cancer Research UK Gurdon Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 1QN, UK; Department of Physiology, Development and Neuroscience, Downing Street, University of Cambridge, Cambridge CB2 3EG, UK; Wellcome Trust-Medical Research Council Stem Cell Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 3EG, UK.

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Related in: MedlinePlus