Study of the expression of a pluripotency-associated Long non coding RNA in Embryonic germ cells

Abstract

Although a wealth of information currently exists for genetic networks involved in pluripotency of mouse embryonic stem cells mESCs, less is known about mouse embryonic germ cells (mEGCs). Especially To date more studies have focused almost on protein-coding transcripts. However, recent transcriptome analyses show that the mammalian genome contains thousands of long noncoding RNAs (LncRNAs), many of which appear to be expressed in a developmentally regulated manner. The purpose of this study is to identify other key regulators of pluripotency in mEGCs.<br><br><br><br>P19 cells were cultured in α-MEM supplemented with 10 % heat-inactivated fetal bovin serum. RNA was extracted from P19 cells in exponential phase. Quality and quantity of RNA was examined by gel electrophoresis and Nano drop. To study the expression of LincRNA1283(large intergenic non coding RNA) at RNA level, at first we found the sequences of this LincRNA in papers and designed specific primer by Primer premier software. Specific cDNA was synthesized by revers primer and hemi nested PCR was done. To confirm amplified PCR product, it was sequenced.<br><br> Hemi nested PCR for lincRNA 1283 produced a fragment of 285 bp size. Its sequence was also confirmed by sequencing. As a result in this study, we showed expression of lincRNA 1283 at mRNA level in P19 cell as pluripotent cell.