Suppression of PRPF4 regulates pluripotency, proliferation, and differentiation in mouse embryonic stem cells

Cell Biochem Funct. 2019 Dec;37(8):608-617. doi: 10.1002/cbf.3437. Epub 2019 Sep 10.

Abstract

Mouse embryonic stem cells (mESCs) are characterized by their self-renewal and pluripotency and are capable of differentiating into all three germ layers. For this reason, mESCs are considered a very important model for stem cell research and clinical applications in regenerative medicine. The pre-mRNA processing factor 4 (PRPF4) gene is known to have a major effect on pre-mRNA splicing and is also known to affect tissue differentiation during development. In this study, we investigated the effects of PRPF4 knockdown on mESCs. First, we allowed mESCs to differentiate naturally and observed a significant decrease in PRPF4 expression during the differentiation process. We then artificially induced the knockdown of PRPF4 in mESCs and observed the changes in the phenotype. When PRPF4 was knocked down, various genes involved in mESC pluripotency showed significantly decreased expression. In addition, mESC proliferation increased abnormally, accompanied by a significant increase in mESC colony size. The formation of mESC embryoid bodies and teratomas was delayed following PRPF4 knockdown. Based on these results, the reduced expression of PRPF4 affects mESC phenotypes and is a key factor in mESC. SIGNIFICANCE OF THE STUDY: Our results indicate that PRPF4 affects the properties of mESCs. Suppression of PRPF4 resulted in a decrease in pluripotency of mESC and promoted proliferation. In addition, suppression of PRPF4 also resulted in decreased apoptosis. Moreover, the inhibition of PRPF4 reduced the ability to differentiate and formation of teratoma in mESC. Our results demonstrated that PRPF4 is a key factor of controlling mESC abilities.

Keywords: PRPF4; differentiation; embryonic stem cells; pluripotency; self-renewal.

MeSH terms

  • Animals
  • Cell Differentiation*
  • Cell Proliferation*
  • Cells, Cultured
  • Mice
  • Mouse Embryonic Stem Cells / cytology
  • Mouse Embryonic Stem Cells / metabolism
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Ribonucleoprotein, U4-U6 Small Nuclear / antagonists & inhibitors
  • Ribonucleoprotein, U4-U6 Small Nuclear / genetics
  • Ribonucleoprotein, U4-U6 Small Nuclear / metabolism*
  • Teratoma / genetics
  • Teratoma / pathology

Substances

  • RNA, Small Interfering
  • Ribonucleoprotein, U4-U6 Small Nuclear