Down-regulate Nanog Expression in Embryonic Stem Cells Using RNA Interference

Abstract: Embryonic stem cells are derived from inner cell mass of blastocyst. Embryonic stem cells have two defining properties， self-renewal and pluripotency， which make them very attractive in clinic. Understanding of the mechanism involved in the pluripotency is essential for achieving these goals. Nanog plays a crucial role in the maintenance of pluripotency. In order to investigate Nanog mechanisms in embryonic stem cells pluripotency， we used RNA interference to specifically down-regulate Nanog expression in embryonic stem cells and investigated the change of J1 cells following with Nanog down-regulation. Here， we designed four siRNAs on the basis of Nanog sequence. Then we analyzed Nanog expression using RT-PCR， Real-Time PCR， and Western blot after these siRNAs were transfected in J1 cells. We observed that Nanog-P1 significantly down-regulated Nanog expression in J1 cells， the precise interference rate of Nanog-P1 was up to 90%. Following Nanog expression was silenced， utf1 (a pluripotency marker of ES cells) expression was decreased but gata6 (a differentiation related gene) was up-regulated. Our experiments showed that Nanog played an important role in ES cells pluripotency.

CSTR: 32200.14.cjcb.2009.01.0020