The Mouse siGENOME RTF Ubiquitin Conjugation 3 siRNA library contains RING finger and RING finger-like E3 Ligases. These enzymes form a site of interaction with E2s through a platform generated as a consequence of coordination of two zinc ions. U-box E3s conform like RING finger proteins, except that they do so through salt bridges and other interactions that do not involve coordination of zinc. A20-like proteins, also included in this set, represent a class of E3 ligases, which contain a single coordination site for zinc.
RTF siRNA libraries are provided as multiple single-use plate sets – just rehydrate, and add cells. This unique pre-plated format reduces hands-on time for faster screening results.
siGENOME siRNAs are designed with the proprietary SMARTselection design algorithm for high-efficiency, guaranteed silencing. They also incorporate rational strand bias with application of ON-TARGET modifications to optimize antisense strand loading into RISC for effective target knockdown.
- Six ready-to-use 96-well plate sets provided for two triplicate screens
- Pre-plated, validated RNAi controls included
- No aliquoting necessary, just resuspend and add cells
- siRNA reagents provided in clear plates at 6.25 pmol per well (50 nM final screening concentration)
- Black or white clear-bottom plates available to support assays involving fluorescent or luminescent detection
|Well||Pre-plated control||Control Catalog No.|
|A1||siGENOME Non-targeting siRNA #2||D-001210-02|
|B1||siGENOME Non-targeting siRNA #3||D-001210-03|
|C1||siGENOME Non-targeting siRNA #4||D-001210-04|
|D1||siGENOME Non-targeting siRNA #5||D-001210-05|
|E1||siGENOME Non-targeting pool #2||D-001206-14|
|F1||siGENOME Cyclophilin B control pool (H, M, R) Control siRNA||D-001136-01|
DharmaFECT transfection reagents are highly recommended for use with RTF libraries and should be purchased separately. Refer to the DharmaFECT Cell Type Guide to find the appropriate formulation for your cell type.
For a complete list of target genes in this siRNA Library, please contact Technical Support or your local Sales Representative.
For a thorough investigation of the ubiquitin pathway, you may also consider our three Ubiquitin Conjugation libraries:
Our siRNA knockdown guarantee
siGENOME and ON-TARGETplus siRNA reagents (SMARTpool and three of four individual siRNAs) are guaranteed to silence target gene expression by at least 75% at the mRNA level when demonstrated to have been used under optimal delivery conditions (confirmed using validated positive control and measured at the mRNA level 24 to 48 hours after transfection using 100 nM siRNA).
Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25 nM working concentration.
Plate layout of siGENOME RTF siRNA Libraries
Figure 1. | Validated control siRNAs and pools are pre-dispensed into column 1 of each RTF Library plate, providing a consistent baseline for screening and assay efficiency.
Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries
Figure 2.| Reverse Transfection Format is highly effective across cell lines
Figure 3. | Reverse transfection Format was used to assess control gene silencing (Cyclophilin B; blue bars) and viability (yellow dots) across eight cell lines under optimized conditions. In all cases, effective target gene knockdown was achieved with low cytotoxicity.
Publications using RTF Libraries
- T. Sorkina, M. Miranda, K. R. Dionne, B. R. Hoover, N. R. Zahniser, A. Sorkin, RNA interference screen reveals an essential role of Nedd4-2 in dopamine transporter ubiquitination and endocytosis. J Neurosci. 26(31), 8195-205 (2006).
- P. Monteiro, D. Gilot, E. Le Ferrec, C. Rauch, D. Lagadic-Gossmann, O. Fardel, Dioxin-mediated up-regulation of aryl hydrocarbon receptor target genes is dependent on the calcium/calmodulin/CaMKIalpha pathway. Mol Pharmacol. 73(3), 769-77 (Epub 18 December 2007, March 2008).
- A. A. Kolokoltsov, D. Deniger, E. H. Fleming, N. J. Roberts Jr, J. M. Karpilow, R. A. Davey, Small interfering RNA profiling reveals key role of clathrin-mediated endocytosis and early endosome formation for infection by respiratory syncytial virus. J Virol. 81(14), 7786-800 (Epub 9 May 2007, July 2007).
- K. M. Hussain, K. L. Leong, M. M. Ng, J. J. Chu, The essential role of clathrin-mediated endocytosis in the infectious entry of human enterovirus 71. J Biol Chem. 286(1), 309-321 (Epub 18 October 2010, 7 January 2011).
General Screening References
- B. D. Parsons, A. Schindler, D. H. Evans, E. Foley, A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay. PLoS One. 4(12), e8471 (2009).
- M. Jiang, R. Instrell, B. Saunders, H. Berven, M. Howell, Tales from an academic RNAi screening facility; FAQs. Brief Funct. Genomics. 10(4), 227-237 (2011). [doi: 10.1093/bfgp/elr016]
The most broadly applicable DharmaFECT formulation for optimal siRNA or microRNA transfection into a wide range of cell types for successful RNAi experiments