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Predesigned siRNA

MISSION® Predesigned siRNA were created using the proprietary Rosetta Inpharmatics siRNA Design algorithm in an exclusive partnership with Merck & Co. The Rosetta siRNA Design Algorithm utilizes Position-Specific Scoring Matrices and knowledge of the seed region to predict the most specific and effective sequences for your target genes. The algorithm’s rules were developed utilizing empirical data collected from gene silencing experiments carried out over three years.

Product Benefits

  • Best-in-class, guaranteed gene silencing
  • Efficient knockdown of low abundance messages
  • Simplified transfection optimization with 11 Positive Control siRNA
  • Distinguish sequence-specific silencing from non–specific effects with 8 negative control siRNA
  • Hundreds of functionally-validated predesigned siRNA

Product Features

  • Species: Human, Mouse & Rat
  • Quantities: 2 (10 nmol), 5 (25 nmol) & 10 (50 nmol) OD
  • Purification: Desalt or HPLC
  • Sequence Form: 21mer duplexes with dTdT overhangs
  • Quality Control: 100% mass spectrometry*
  • Format: Supplied dry in tubes

*Depending on manufacturing site, PAGE may be used to assess siRNA duplexes.

Predesigned Guarantee

At least two of three purchased MISSION® Predesigned siRNA targeting the same gene will reduce target mRNA levels by 75%, in cultured cells, when transfected at a concentration of ≥30 nM. If two of the siRNA do not knock down the target gene by 75%, we will provide three additional siRNA for that gene, free of charge. If there are no more siRNA for that gene or all siRNA fail to knock down the target gene by 75%, we will refund the purchase price.
Receipt of appropriate supporting data for transfection efficiency is required for the guarantee to apply. Appropriate supporting data for transfection efficiency would include qPCR data comparing target mRNA levels of a MISSION Positive Control siRNA (GAPDH, MAPK1, TP53, etc.), transfected at ≥30 nM, to an appropriate negative control (such as mock transfection, a scrambled siRNA sequence, or MISSION Universal Negative Control siRNA), demonstrating knockdown of the target mRNA of 75%.
Due to the variability of antibodies and protein half-lives, we are unable to accept data from protein-based detection methods.

 Product Pools

A popular pooled format is 4 duplexes at 5 nmol each combined into one tube (20 nmol pooled) plus the exact same 4 duplexes also at 5 nmol each in separate tubes (another 20 nmol individual). However, our sophisticated liquid handlers allow for a wide range of other possibilities. For feasibility review of your specific needs, contact sirnarequest@sial.com.

Validated siRNA

Many common gene targets have been validated for ≥75% mRNA knockdown (Figure 1 for example data and the table for a list of commonly-ordered, validated siRNA by gene symbol). Validated siRNA are suitable for transfection optimization and as positive controls.

HeLa cells transfected with predesigned siRNA at a concentration of 30 nM.

Figure 1.HeLa cells transfected with predesigned siRNA at a concentration of 30 nM. The percentage remaining gene expression levels were measured via qPCR 48 hours after transfection (relative to mock). Data represents the mean of four biological replicates.

Materials
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Select Citations

1.
Yang X, Sierant M, Janicka M, Peczek L, Martinez C, Hassell T, Li N, Li X, Wang T, Nawrot B. 2012. Gene Silencing Activity of siRNA Molecules Containing Phosphorodithioate Substitutions. ACS Chem. Biol.. 7(7):1214-1220. https://doi.org/10.1021/cb300078e
2.
Salma J, McDermott JC. 2012. Suppression of a MEF2-KLF6 Survival Pathway by PKA Signaling Promotes Apoptosis in Embryonic Hippocampal Neurons. Journal of Neuroscience. 32(8):2790-2803. https://doi.org/10.1523/jneurosci.3609-11.2012
3.
Gilot D, Le Meur N, Giudicelli F, Le Vée M, Lagadic-Gossmann D, Théret N, Fardel O. RNAi-Based Screening Identifies Kinases Interfering with Dioxin-Mediated Up-Regulation of CYP1A1 Activity. PLoS ONE. 6(3):e18261. https://doi.org/10.1371/journal.pone.0018261
4.
Raab M, Kappel S, Krämer A, Sanhaji M, Matthess Y, Kurunci-Csacsko E, Calzada-Wack J, Rathkolb B, Rozman J, Adler T, et al. 2011. Toxicity modelling of Plk1-targeted therapies in genetically engineered mice and cultured primary mammalian cells. Nat Commun. 2(1): https://doi.org/10.1038/ncomms1395
5.
Chia KM, Liu J, Francis GD, Naderi A. 2011. A Feedback Loop between Androgen Receptor and ERK Signaling in Estrogen Receptor-Negative Breast Cancer. Neoplasia. 13(2):154-166. https://doi.org/10.1593/neo.101324
6.
Ramachandran V, Arumugam T, Langley R, Hwang RF, Vivas-Mejia P, Sood AK, Lopez-Berestein G, Logsdon CD. The ADMR Receptor Mediates the Effects of Adrenomedullin on Pancreatic Cancer Cells and on Cells of the Tumor Microenvironment. PLoS ONE. 4(10):e7502. https://doi.org/10.1371/journal.pone.0007502
7.
Santra MK, Wajapeyee N, Green MR. 2009. F-box protein FBXO31 mediates cyclin D1 degradation to induce G1 arrest after DNA damage. Nature. 459(7247):722-725. https://doi.org/10.1038/nature08011
8.
Meng W, Mushika Y, Ichii T, Takeichi M. 2008. Anchorage of Microtubule Minus Ends to Adherens Junctions Regulates Epithelial Cell-Cell Contacts. Cell. 135(5):948-959. https://doi.org/10.1016/j.cell.2008.09.040
9.
Matsubara T, Kida K, Yamaguchi A, Hata K, Ichida F, Meguro H, Aburatani H, Nishimura R, Yoneda T. 2008. BMP2 Regulates Osterix through Msx2 and Runx2 during Osteoblast Differentiation. J. Biol. Chem.. 283(43):29119-29125. https://doi.org/10.1074/jbc.m801774200
10.
Zhou H, Xu M, Huang Q, Gates AT, Zhang XD, Castle JC, Stec E, Ferrer M, Strulovici B, Hazuda DJ, et al. 2008. Genome-Scale RNAi Screen for Host Factors Required for HIV Replication. Cell Host & Microbe. 4(5):495-504. https://doi.org/10.1016/j.chom.2008.10.004
11.
Espeseth AS, Huang Q, Gates A, Xu M, Yu Y, Simon AJ, Shi X, Zhang X, Hodor P, Stone DJ, et al. 2006. A genome wide analysis of ubiquitin ligases in APP processing identifies a novel regulator of BACE1 mRNA levels. Molecular and Cellular Neuroscience. 33(3):227-235. https://doi.org/10.1016/j.mcn.2006.07.003
12.
Bartz SR, Zhang Z, Burchard J, Imakura M, Martin M, Palmieri A, Needham R, Guo J, Gordon M, Chung N, et al. 2006. Small Interfering RNA Screens Reveal Enhanced Cisplatin Cytotoxicity in Tumor Cells Having both BRCA Network and TP53 Disruptions. MCB. 26(24):9377-9386. https://doi.org/10.1128/mcb.01229-06
13.
Majercak J, Ray WJ, Espeseth A, Simon A, Shi X, Wolffe C, Getty K, Marine S, Stec E, Ferrer M, et al. 2006. LRRTM3 promotes processing of amyloid-precursor protein by BACE1 and is a positional candidate gene for late-onset Alzheimer's disease. Proceedings of the National Academy of Sciences. 103(47):17967-17972. https://doi.org/10.1073/pnas.0605461103

If additional help is needed, please consult our technical services group at oligotechserv@sial.com.

MISSION is a trademark of Merck KGaA, Darmstadt, Germany and/or its affiliates. Label License.

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