Double-stranded RNA (dsRNA) is not only a hallmark of viral infections but also a key molecule in RNA interference (RNAi)-mediated gene silencing at the mRNA level.
Large amounts of >200 bp dsRNA (> 2 mg/ml transcription reaction) are efficiently produced using the HighYield T7 RNAi Kit via T7 RNA Polymerase-mediated in vitro transcription (Fig. 1). The resulting dsRNA can subsequently be used for RNA interference (RNAi) applications by direct administration e.g. on plant tissues[1] or as template for enzymatic digestion to short interfering RNA (siRNA)[2-4].
Figure 1: Similar results with Jena Bioscience’s HighYield T7 RNAi Kit and the T7 RiboMAX™ Express RNAi System.
A) dsRNA is synthesized in a 3 step procedure.
B) Similar transcription efficiency of Jena Bioscience’s HighYield T7 RNAi Kit and T7 RiboMAX™ Express RNAi System. In vitro transcription has been performed according to the manufacturer protocol using 1 µg DNA template 1. Transcription efficiency was analyzed by agarose gelelectrophoresis (equal volumes of in vitro transcription reactions). JBS: HighYield T7 RNAi Kit, Promega: T7 RiboMAX™ Express RNAi System
C) Competitive pricing of Jena Bioscience’s HighYield T7 RNAi Kit.
[1] Voloudakis et al. (2015) Efficient Double-Stranded RNA Production Methods for Utilization in Plant Virus Control. Plant Virology Protocols. Methods in Molecular Biology 1236:1.
[2] Sun et al. (2018) A Simple and Cost-Effective Approach for in vitro production of sliced siRNAs as potent triggers for RNAi. Molecular Therapy Nucleic Acids 8:345.
[3] Kittler et al. (2005) Production of endoribonuclease-prepared short interfering RNAs for gene silencing in mammalian cells. Nature Methods 2 (10): 779.
[4] Buchholz et al. (2006) Enzymatically prepared RNAi libraries. Nature Methods 3 (9):696.
