Stem-Loop RT-PCR Quantification of siRNAs In Vitro and In Vivo

被引：39

作者：

Cheng, Angie ^{[1
]}

Li, Mu ^{[1
]}

Liang, Yu ^{[2
]}

Wang, Yu ^{[2
]}

Wong, Linda ^{[2
]}

Chen, Caifu ^{[2
]}

Vlassov, Alexander V. ^{[1
]}

Magdaleno, Susan ^{[1
]}

机构：

[1] Life Technol, Austin, TX 78744 USA

[2] Life Technol, Foster City, CA USA

来源：

OLIGONUCLEOTIDES | 2009年 / 19卷 / 02期

关键词：

GENE-EXPRESSION; DELIVERY; RNAI; DNA;

D O I：

10.1089/oli.2008.0176

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

RNA interference (RNAi) is a mechanism in which the introduction of small interfering RNAs (siRNAs) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. Applications of RNAi include gene function and pathway analysis, target identification and validation, and therapeutics. There is a need to develop reliable and easy-to-use assays to evaluate siRNA delivery efficiency and distribution, study pathways, and stability of siRNAs in cells (posttransfection) and in animals (postinjection). We have leveraged the Applied Biosystems TaqMan (R)-based stem-loop RT-PCR technology, originally developed for quantification of endogenous microRNAs in cells, to fulfill these needs. The application protocols developed enable robust quantification of siRNA, including chemically modified siRNA molecules, in vitro and in vivo.

引用

收藏

页码：203 / 208

页数：6

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