The Mouse siGENOME Cell Cycle Regulation siRNA Library includes siRNA reagents targeting genes whose functions are crucial for controlling cell division. Of particular importance to the progression of the cell cycle are the cyclin-dependent kinases, or CDKs. Other protein families known to regulate the cell cycle include the members of the CIP/KIP family and the INK4 family of cell cycle inhibitors, members of the retinoblastoma protein family and DNA replication factors, such as the cell division cycle proteins (CDCs).
siGENOME siRNA reagents are globally recognized and trusted for highly effective performance in RNAi screening applications (see Supporting Data tab). Our expertise in siRNA design, experimental optimization, and siRNA screening strategies provide end-to-end support of your screening efforts.
- Guaranteed target knockdown (see Specifications tab)
- Antisense strand loading into RISC ensured by thermodynamic analysis and selective application of a sense strand-blocking modification (ON-TARGET)
- Available as SMARTpool or Set of 4 siRNA reagents arrayed in 96-well plates
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 nM siRNA).
Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25 nM working concentration.
Screening with siGENOME SMARTpool siRNA reagents provide effective target silencing
The effect of silencing ARPC1B on cell migration was studied in a breast cancer cell line. A monolayer of cells was uniformly scraped, and the rate of cell migration to close the scrape (wound healing) was evaluated. Both unmodified and ON-TARGETplus siRNA reagents induced potent target knockdown. Inconsistent phenotypes due to off-target effects (red outline) were observed for cells transfected with unmodified individual siRNAs.
The unmodified SMARTpool improved the false phenotype considerably while the ON-TARGETplus SMARTpool significantly reduced off-target effects to produce a consistent phenotype. In collaboration with Kaylene Simpson, Laura Selfors, and Joan Brugge, Harvard Medical School.
Unnecessary sense-strand inactivation can increase off-target activity
Unmodified and sense strand-inactivated siRNAs were used to target five genes. In four cases, off-targets were increased due to enhanced RISC loading of the antisense strand when the sense strand was modified. The unmodified siRNAs have natural guide-strand loading characteristics. All siRNAs had comparable silencing potency. Data shown represents genes down-regulated by twofold or more. HEK293 cells were transfected with 100 nM siRNA using 0.2 μL of DharmaFECT 1. Data was analyzed at 24 hours by genomewide microarray analysis (Agilent).
Why not modify ALL siGENOME siRNAs to ensure proper strand loading? It has been demonstrated by Dharmacon scientists and others that forcing antisense (guide) strand entry into RISC may actually INCREASE off-targets due to increased loading of the guide strand and resulting off-target activity by its seed region. siGENOME siRNAs are designed with thermodynamic properties to naturally facilitate guide strand entry to RISC, which has been demonstrated to correlate with functionality. However, in cases where a high-scoring siGENOME siRNA does not possess ideal strand-loading characteristics, a sense (passenger) strand-inhibiting chemical modification (ON-TARGET) is utilized to promote guide strand entry. Approximately 20% of siGENOME siRNAs carry the ON-TARGET modification.
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- J. A. Smith et al., Genome-wide siRNA screen identifies SMCX, EP400, and Brd4 as E2-dependent regulators of human papillomavirus oncogene expression. Proc. Natl. Acad. Sci. U S A. 107(8), 3752-3757. (Epub 2 February 2010, 23 February 2010).
- G. Hu et al., A genome-wide RNAi screen identifies a new transcriptional module required for self-renewal. Genes Dev. 23(7), 837-848 (1 April 2009).
- R. D. Paulsen et al., A genome-wide siRNA screen reveals diverse cellular processes and pathways that mediate genome stability. Mol Cell. 35(2), 228-239 (31 July 2009).
- M. Tsui et al., An intermittent live cell imaging screen for siRNA enhancers and suppressors of a kinesin-5 inhibitor. PLoS One. 4(10), e7339 (5 October 2009).
- A. L. Brass et al., Identification of host proteins required for HIV infection through a functional genomic screen. Science. 319(5865), 921-926. (Epub 10 January 2008, 15 February 2008).
- C. Swanton et al., Regulators of mitotic arrest and ceramide metabolism are determinants of sensitivity to paclitaxel and other chemotherapeutic drugs. Cancer Cell. 11(6), 498-512 (11 June 2007).
- A. W. Whitehurst et al., Synthetic lethal screen identification of chemosensitizer loci in cancer cells. Nature. 446(7137), 815-819 (12 April 2007).
Concentrated buffer solution recommended for resuspension and long-term storage of any short, double-strand, or single-strand synthetic RNA molecule. Dilute with RNase-free water prior to use.
Molecular grade water for dilution of 5x siRNA Buffer or resuspension of RNA. RNase-free to prevent degradation of RNA reagents and oligonucleotides.
An aliquot of each of the four DharmaFECT formulations for siRNA/microRNA transfection optimization studies