一种用于哺乳动物细胞RNA干扰的通用表达载体的设计与应用

Design and application of a versatile expression vector for RNAi in mammalian cells.

作者信息

Lin Xia, Feng Xin-Hua

机构信息

Michael E. DeBakey Department of Surgery, and Department of Molecular & Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA.

出版信息

J RNAi Gene Silencing. 2005 Jul 28;1(1):38-43.

PMID:19771203

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2737199/

Abstract

Here we report a versatile mammalian expression vector called pRIGHT11 for production of small interfering RNA (siRNA) in cells. This vector uses opposing eukaryotic RNA polymerase III promoters U6 and H1 to drive the expression of short siRNA. We have demonstrated the effectiveness of pRIGHT11-generated siRNA in sequence-specific inhibition of transfected reporter genes and endogenous genes. Furthermore, this retrovirus-based vector can carry a random library of siRNA, and thus can be applied to rapid screening of novel genes involved in specific cellular responses.

摘要

在此，我们报道了一种多功能的哺乳动物表达载体pRIGHT11，用于在细胞中产生小干扰RNA（siRNA）。该载体利用反向的真核RNA聚合酶III启动子U6和H1来驱动短siRNA的表达。我们已经证明了pRIGHT11产生的siRNA在对转染的报告基因和内源基因进行序列特异性抑制方面的有效性。此外，这种基于逆转录病毒的载体可以携带siRNA随机文库，因此可应用于快速筛选参与特定细胞反应的新基因。