Gene Silencing Delivery Methods: Lipid-Mediated and Electroporation Transfection Protocols

Methods Mol Biol. 2016:1434:139-51. doi: 10.1007/978-1-4939-3652-6_10.

Abstract

The RNA interference (RNAi) plays an important role in regulation of gene expression. It is a mechanism used by many organisms to silence the expression of genes that control different processes in the cell. The double strand (ds) RNA molecule inhibits gene expression of a targeted gene with high specificity and selectivity.Different types of small ribonucleic acid molecules, microRNA (miRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), and the piwi RNA (piRNA) are involved in the RNA interference. RNAi is a relevant research tool in cell cultures and in vivo experiments because synthetic dsRNA introduced into cells can selectively silence specific target genes.Here, we describe a general guide for gene silencing mediated by siRNA, focusing on the most used delivery methods: lipid-mediated and electroporation transfection.

Keywords: Electroporation transfection; Lipid-mediated transfection; mRNA regulation RNA interference; miRNA; shRNA; siRNA.

MeSH terms

  • Cell Adhesion
  • Cells, Cultured
  • Electroporation / methods*
  • HEK293 Cells
  • Humans
  • Lipids / chemistry*
  • RNA Interference
  • Ribonuclease III / metabolism
  • Transfection / methods*

Substances

  • Lipids
  • Lipofectamine
  • Ribonuclease III