GSE100285
Trophectoderm specific function of Atypical Protein Kinase C iota (PKCi) is essential for post-implantation mammalian development.
Atypical protein kinase C iota (Prkci) knockout mice are embryonic lethal at gestation day 9.5 but the underlying molecular mechanisms is not known. Here , using Prkci knockout mouse model , we show that trophectoderm specific function of PKCi is essential for post-implantation mammalian development. We observed that PKCi is expressed predominantly in the trophectoderm lineage and developing placenta region in post-implantation mouse embryos. Prkci knockout or homozygous embryos show severe defect in placenta formation compared to wildtype and heterozygous embryos.  Using RNA seq analysis in PKCi knockdown mouse trophoblast stem cells, we identified certain genes which are significantly upregulated upon PKCi depletion , one among them is Bone Morphogenetic protein 13 (Bmp13). Our study shows that PKCi is one of key players involved in proper development of the placenta, loss of which might be one of the reasons for early pregnancy failure. Overall design: Mouse trophoblast stem cells were cultured using MEF conditioned media along with FGF4 and Heparin. Short hairpin RNA against Prkci mRNA was cloned in PLKO1 vector and a scrambled control was also used. Lentiviral supernatants were obtained from HEK 293T cells and were used to transduce TS cells described before. Transduced cells were selected using 1µg/ml puromycin. Knockdown efficiency was validated by real-time PCR analysis and western blotting. Whole cell-lysates were used to prepare RNA using Qiagen RNeasy Mini Kit with on column DNase digestion. RNA-seq analysis were performed using Illumina HiSeq2500 platform.