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Summary: Overview: RNA interference (RNAi) by double stranded RNA (dsRNAs) molecules of approximately 20-25 nucleotides termed short interfering (siRNAs) is a powerful method for preventing the expression of a particular gene. The dsRNA dominantly silences gene expression in a sequence-specific manner by causing the corresponding endogenous mRNA to be degraded. The technique was first developed in Caenorhabditis elegans, and was rapidly applied to a wide range of organisms. Methods for expressing siRNAs in cells in culture and in vivo using viral vectors, and for transfecting cells with synthetic siRNAs, have been developed and are being used to establish the functions of specific proteins in various cell types and organisms. For example, chemically synthesized or in vitro transcribed siRNAs can be transfected into cells, injected into mice, or introduced into plants. siRNAs can also be expressed endogenously from siRNA expression vectors or PCR products in cells or in transgenic animals.
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Protocols
- dsRNA Synthesis Protocol (Drosophila RNAi Screening Center at Harvard Medical School)
Generating dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends.
http://flyrnai.org/DRSC-PRS.html
Added: Wed May 12 2004, Hits: 2592, Reviews: 0 Write review Cached - The siRNA User Guide (Tom Tuschl, Rockefeller University )
Offers guidance and tips on the following topics: 1) Selection of siRNA duplexes from the target mRNA sequence; 2)Preparation of siRNA duplexes; 3)Useful siRNA reagent combinations; 4)Transfection of siRNA duplexes.
http://www.rockefeller.edu/labheads/tuschl/sirna.h...
Added: Fri Jun 20 2003, Hits: 6292, Reviews: 1 Read reviews Write review Cached -
Bacterial mediated RNAi
(Dr. Ian Chin-Sang)
Bacterial mediated RNAi using HT115 competent cell cultures.
http://research.abl.es/methods/1427/
Added: Fri Feb 17 2012, Hits: 1045, Reviews: 0 Write review -
DsRNAi Experiments in Drosophila Cell
(Laboratory of Jack Dixon, UCSD)
RNAi takes advantage of the unique ability of double-stranded RNA (dsRNA) molecules to induce posttranscriptional gene silencing in a highly specific manner. This silencing is efficacious and long-lived, as it is passed to subsequent generations in insect cell culture. To date, all Drosophila cell lines tested (S2, KC, BG2-C6, and Shi) respond to dsRNAs by ablating expression of the target protein. Furthermore, all dsRNAs tested (more than 15) have been effective at silencing the target gene.
http://dixonlab.biochem.med.umich.edu/RNAiExp.html
Added: Fri May 09 2003, Hits: 1201, Reviews: 0 Write review Cached -
MicroRNA and siRNA Cloning Protocol
(Bartel Lab, MIT)
Detailed procedures on oligo design, synthesis, purification and cloning.
http://web.wi.mit.edu/bartel/pub/protocols/miRNAcl...
Added: Sat May 24 2003, Hits: 3445, Reviews: 1 Read reviews Write review -
RNA Interference (RNAi) Protocol
(Forler D, et al.)
This method uses the Promega Ribomax Large Scale RNA Production System T7 to produce dsRNAs and RNAi is conducted in Drosophila Schneider cells.
http://www.mann.embl-heidelberg.de/GroupPages/Page...
Added: Thu May 08 2003, Hits: 764, Reviews: 0 Write review Cached -
RNA Interference (RNAi) Resources
(Ambion)
If you are new to RNAi, here you can find some good stuff related to RNAi, such as the mechanism of RNAi, how to design SiRNA.
http://www.ambion.com/techlib/resources/RNAi/
Added: Thu May 08 2003, Hits: 2869, Reviews: 0 Write review Cached
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