- Easily and economically generates functional siRNAs
- No guesswork: An siRNAs mix has a better chance of success than a single siRNA design
- No wasted time and money due to failed siRNA designs
- More regions of the genes can be screened for silencing
- Provided with the proven GeneSilencer siRNA Transfection Reagent
The Dicer siRNA Generation Kit allows easy and cost-effective generation of a large number of small interfering RNAs (siRNAs) from full-length target genes.
The Dicer siRNA Generation Kit allows easy and cost-effective generation of a large number of small interfering RNAs (siRNAs) from full-length target genes (1).
siRNAs are 21- to 23-nucleotide RNA molecules that can cause targeted gene silencing in mammalian cells through a process known as RNA interference (2,3,4).
In nature, siRNAs are generated by ribonuclease III cleavage of longer double stranded RNAs (dsRNAs).
When dsRNAs are transfected directly into mammalian cells, they activate the interferon system and provoke non-specific gene suppression and cytotoxic response (2).
siRNAs have proven to be effective at specifically silencing gene expression without causing any interferon response.
The Dicer siRNA Generation Kit mimics the natural RNA interference process by using recombinant human dicer enzyme, a double-stranded RNA-specific endonuclease, to cleave in vitro transcribed dsRNA templates into a pool of 22 bp siRNAs (Figure 1, 2).
Figure 1. The Dicer siRNA Generation Kit contains everything that is required for preparing double stranded RNA, RNA cleavage, siRNA cleanup and transfection. Both the Recombinant Dicer Enzyme and the RNA Purification Columns are also available separately.
Figure 2. Hela cells were transiently tranfected with 1ug of gWIZ/GFP. When co-transfected with 500ug of Diced GFP siRNA, GFP expression was efficiently suppressed. Cells were imaged 48 hours post tranfection. (GFP: green, left panels. DAPI: blue, right panels)
1. Myers, J.W. et al. (2003) Nature Biotechnology 21: 324-328.
2. Elbashir, S.M. et al. (2001) Nature 411: 494-498.
3. Caplen, N.J. et al. (2001) Proc Natl Acad Sci USA 98: 9742-9747.
4. Sharp, P.A. (2001) Genes and Development 15: 485-490.
Sufficient material is provided for transcribing, cleaving, and transfecting siRNAs for 50 transfection experiments in 24-well plates and with up to 5 different genes.
The Dicer siRNA Generation Kit is shipped frozen. Both the TurboScript T7 Transcription Kit and the recombinant human dicer enzyme should be stored at -20oC upon receipt. If stored properly, all reagents are stable for 6 months.