Stem-loop RT-PCR quantification of siRNAs in vitro and in vivo.
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-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.
Cheng
Angie
A
Life Technologies, Austin, TX 78744, USA.
Li
Mu
M
Liang
Yu
Y
Wang
Yu
Y
Wong
Linda
L
Chen
Caifu
C
Vlassov
Alexander V
AV
Magdaleno
Susan
S
D000818:Animals; D045744:Cell Line, Tumor; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000032:analysis* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014162:Transfection
D034741:RNA, Small Interfering