Conditional Suppression of Cellular Genes: Lentivirus Vector-Mediated Drug-Inducible RNA Interference

TLDR

RNA interference is a powerful tool for downregulating specific genes in cells and animals, opening new possibilities in developmental genetics and molecular therapeutics. The study presents a method that expands RNA interference by enabling conditional suppression of genes in mammalian cells. The method employs a lentiviral vector to place a polymerase III promoter‑driven siRNA cassette under doxycycline‑controlled transcriptional repression. The system achieves highly efficient, fully drug‑inducible knockdown of cellular genes and, because it uses lentiviral delivery, offers broad applicability in vitro, in vivo, and for transgenic animal production.

Abstract

ABSTRACT RNA interference has emerged as a powerful technique to downregulate the expression of specific genes in cells and in animals, thus opening new perspectives in fields ranging from developmental genetics to molecular therapeutics. Here, we describe a method that significantly expands the potential of RNA interference by permitting the conditional suppression of genes in mammalian cells. Within a lentivirus vector background, we subjected the polymerase III promoter-dependent production of small interfering RNAs to doxycycline-controllable transcriptional repression. The resulting system can achieve the highly efficient and completely drug-inducible knockdown of cellular genes. As lentivirus vectors can stably transduce a wide variety of targets both in vitro and in vivo and can be used to generate transgenic animals, the present system should have broad applications.