A Practical Protocol for the Conditional Depletion of Rho Isoforms in Human Embryonic Stem Cells

Methods Mol Biol. 2018:1821:283-295. doi: 10.1007/978-1-4939-8612-5_20.

Abstract

Human pluripotent stem cells indefinitely proliferate and survive in culture while retaining genomic integrity, providing a unique opportunity to study human molecular biology. Here, we introduced an RNA interference-based protocol of inducible gene silencing in human embryonic stem cells, which has several advantages in handling simplicity/convenience, cost/time performance, and applicability. Using this method, we had succeeded to elucidate the isoform-unique roles of Rho-family small GTPases in human embryonic stem cells.

Keywords: Human embryonic stem cells; Inducible RNAi; Lentivirus; Rho small GTPase.

MeSH terms

  • Gene Silencing*
  • HEK293 Cells
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / enzymology*
  • Humans
  • Lentivirus*
  • RNA Interference*
  • Transduction, Genetic / methods*
  • rho GTP-Binding Proteins* / antagonists & inhibitors
  • rho GTP-Binding Proteins* / genetics
  • rho GTP-Binding Proteins* / metabolism

Substances

  • rho GTP-Binding Proteins