The ON-TARGETplus Human Epigenetics siRNA Library targets enzyme classes characterized to play a role in epigenetic mechanisms (see Reference 1). These inheritable factors that regulate genetic expression are of growing importance to scientific understanding of disease progression and trait inheritance. Modification of histones, such as methylation or acetylation, plays a key role in epigenetics as the extent to which DNA is wrapped around these protein complexes will alter the availability of genes in those DNA segments to be activated. The Epigenetics library targets genes which encode histone and DNA modification factors like methyltransferases, acetyltransferases, kinases and deacetylases, in addition to genes with a role in chromatin remodeling and histone ubiquitination.
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.
ON-TARGETplus modifications combine sense strand inactivation with a novel seed-region modification. This patented siRNA approach is the best strategy to prevent off-target effects caused by both the sense and antisense strands while maintaining high silencing potency.
- 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||ON-TARGETplus Non-targeting siRNA #1||D-001810-01|
|B1||ON-TARGETplus Non-targeting siRNA #2||D-001810-02|
|C1||ON-TARGETplus Non-targeting siRNA #3||D-001810-03|
|D1||ON-TARGETplus Non-targeting siRNA #4||D-001810-04|
|E1||ON-TARGETplus Non-targeting pool||D-001810-10|
|F1||ON-TARGETplus Human Cyclophilin B control pool||D-001820-10|
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.
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 µnM siRNA).
Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25 µnM working concentration.
Plate layout of ON-TARGETplus RTF siRNA Libraries
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.
Reverse Transfection Format is highly effective across cell lines
Eight cell lines were transfected with control siRNAs targeting Cyclophilin B using RTF plates. Good cellular viability and effective target silencing were accomplished under optimized conditions. Gene knockdown is normalized to GAPDH expression. Cell plating densities are indicated in parentheses. Gene expression was determined 48 hours posttransfection by branched DNA assay (Panomics Quantigene® Reagent System) (blue bars). Cell viability was determined by alamarBlue® metabolic assay (gold circles).
Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries
Publications using RTF Libraries
- J. Comley. Epigenetics: An emerging target class for drug screening.Drug Discovery World Spring 2011, 40-55 (2011).
- 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]