RNA binding protein PCBP1 regulates hESC pluripotency through modulating mRNA export

Abstract

Objective To investigate the biological function of RNA binding protein polycytosine-binding protein 1 (PCBP1) in the maintenance of pluripotency of human embryonic stem cells (hESC) and to explore the new mechanism of regulating hESC pluripotency. Methods PCBP1 knockdown technology was used to detect the clonogenicity. The alkaline phosphatase staining profile and the expression of pluripotency/differentiation marker genes were detected by qPCR. Meanwhile, the nucleus and cytoplasm of hESC were isolated for RNA sequencing, and the nucleoplasmic changes of RNA generated by PCBP1 knockdown were analyzed by multi-level analysis. Results PCBP1 knockdown significantly inhibited clonogenesis of human embryonic stem cells (P＜0.01) and reduced the RNA level of pluripotency related genes (P＜0.01). The expression of differentiation related genes increased(P＜0.01)that significantly affected the overall nucleoplasmic distribution of RNA in hESC and the nucleoplasmic distribution of mRNA related to embryonic development and cell differentiation. Conclusions PCBP1 regulates hESC pluripotency by regulating the expression of pluripotency/differentiation marker genes and the nucleoplasmic localization of mRNA.

Keywords

• human embryonic stem cell|polycytosine binding protein 1|pluripotency and self-renewal|nuclear and cytoplasm extraction