A System for Stable Expression of Short Interfering RNAs in Mammalian Cells

TLDR

Mammalian genetic studies are limited by the lack of efficient tools for stable loss‑of‑function phenotypes. The authors introduce the pSUPER vector system to generate stable siRNA expression in mammalian cells. The vector directs synthesis of siRNAs, enabling efficient and specific gene knockdown. Stable siRNA expression via pSUPER leads to efficient, specific, and persistent gene silencing, providing a powerful tool for long‑term loss‑of‑function studies in diverse mammalian cell types.

Abstract

Mammalian genetic approaches to study gene function have been hampered by the lack of tools to generate stable loss-of-function phenotypes efficiently. We report here a new vector system, named pSUPER, which directs the synthesis of small interfering RNAs (siRNAs) in mammalian cells. We show that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes. Stable expression of siRNAs using this vector mediates persistent suppression of gene expression, allowing the analysis of loss-of-function phenotypes that develop over longer periods of time. Therefore, the pSUPER vector constitutes a new and powerful system to analyze gene function in a variety of mammalian cell types.