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Transcription profiling of mouse germ cells with conditional Nanog knockdown.

ABSTRACT: Single Cell Microarray
E10.5 genital ridges (TM at E7.5) incubated in 0.5mM EDTA/ PBS for 20 minutes at 37ï¾°C were transferred to 2% BSA/ PBS. PGCs were collected from the genital ridges pierced with fine glass needles. Released PGCs were identified by their morphological characteristics and transferred into lysis buffer with a mouth pipette. PGCs were genotyped with the remaining genital ridges. The amplified cDNA library of each PGC was classified by qPCR-based expression analyses of Nanog, Oct4, and Stella/Dppa3 (Kurimoto et al., 2007). cDNAs were labeled by in vitro transcription (Affymetrix). The cRNAs were hybridized with the GeneChip Mouse Genome 430 2.0 Array (Affymetrix). Data were analyzed using the Microsoft Excel and MeV (multiple experimental viewer) software.

ORGANISM(S): Mus musculus

SUBMITTER: Shinpei Yamaguchi

PROVIDER: E-MEXP-2411 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Conditional knockdown of Nanog induces apoptotic cell death in mouse migrating primordial germ cells.

Yamaguchi Shinpei S Kurimoto Kazuki K Yabuta Yukihiro Y Sasaki Hiroyuki H Nakatsuji Norio N Saitou Mitinori M Tada Takashi T

Development (Cambridge, England) 20091201 23

The pluripotency factor Nanog is expressed in peri-implantation embryos and primordial germ cells (PGCs). Nanog-deficient mouse embryos die soon after implantation. To explore the function of Nanog in germ cells, Nanog RNA was conditionally knocked down in vivo by shRNA. Nanog shRNA transgenic (NRi-Tg) mice were generated through the formation of germline chimeras with NRi-Tg embryonic stem cells. In E12.5 Cre-induced ER-Cre/NRi-Tg and TNAP-Cre/NRi-Tg double-transgenic embryos, the number of alk ...[more]

PMID: 19906868

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