T7 RiboMAX™ Express RNAi System
from
Promega
Description
The T7 RiboMAX™ Express RNAi System(a,b,c) is an in vitro transcription system designed for producing milligram amounts of double-stranded RNA (dsRNA) in a short amount of time. The dsRNA is free of protein and other contaminants and is suitable for use in RNA interference (RNAi) in both mammalian and nonmammalian systems. The T7 RiboMAX™ Express RNAi System can be used to synthesize short interfering RNAs (siRNAs) of 21bp for use in mammalian systems. siRNAs synthesized in vitro have been demonstrated to be as effective as chemically synthesized siRNAs for inducing RNAi in mammalian cells (1,2). Using this system, two complementary RNA strands are synthesized from DNA templates supplied by the user. The resulting RNA strands are annealed after the transcription reaction to form siRNA. The siRNA duplex is then purified and can be introduced into the appropriate mammalian cell line. In addition, the T7 RiboMAX™ Express RNAi System can be used for the synthesis of dsRNA molecules of approximately 200bp or greater, which can be applied to nonmammalian systems. Two complementary RNA strands are synthesized from DNA template (either plasmid or PCR(d) product). The resulting RNA strands are annealed after the transcription reaction to form dsRNA. Any remaining single-stranded RNA and DNA template are removed with a nuclease digestion step. The dsRNA is then purified by isopropanol precipitation and can be introduced into the organism of choice for RNAi applications. The siRNA Target Designer program (www.promega.com/siRNADesigner/) designs oligonucleotides for use with Promega RNA interference systems. The program analyzes input DNA or RNA sequences for regions that fit siRNA design requirements. siRNAs that could target these regions are displayed along with the oligonucleotides required for use with the chosen system.