Human siGENOME RTF - Cell Cycle Regulation

Ideal for labs who want to carry out siRNA screens but lack high-throughput capabilities.


A ready-to-use reverse transfection format RNAi screening library targeting human cell cycle regulation genes. Just resuspend pre-dispensed siRNA and add cells. Optimization plates available.

The siGENOME Cell Cycle Regulation RTF siRNA Library includes SMARTpool 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.

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.

 

Highlights

  • 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
  • SMARTpool 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 controls in ON-TARGETplus RTF Libraries 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

For a diagram of the Optimization Plate layout, see Figure 1.

Experimental considerations

DharmaFECT Cell Culture Reagent (DCCR) is recommended for use with RTF libraries to dilute transfection reagents prior to use. DCCR may be purchased separately or accompanying your RTF Optimization Plates

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.

Gene Targets

For a complete list of target genes in this siRNA Library, please email Technical Support, or call 1-800-235-9880. International customers, please call 303-604-9499 or your local Sales Representative.

  
HazardousNo
Shelf Life12 Months
Shipping ConditionAmbient
Storage Condition4 C

 

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 used under optimal delivery conditions (confirmed using validated positive control and measured at the mRNA level 24 - 48 hours after transfection using 100 nM siRNA).

Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5-25 nM working concentration.

Plate layout of siGENOME RTF siRNA Libraries

Plate layout of siGENOME RTF siRNA Libraries

Plate layout of siGENOME 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

Reverse Transfection Format is highly effective across cell lines

Reverse Transfection Format is highly effective across cell lines

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.


Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries

Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries

Simple four-step protocol for screening with RTF SMARTpool siRNA Libraries


References

  1. Publications using RTF Libraries

    Sorkina T, Miranda M, Dionne KR, Hoover BR, Zahniser NR, Sorkin A. RNA interference screen reveals an essential role of Nedd4-2 in dopamine transporter ubiquitination and endocytosis. J Neurosci. 2006 Aug 2;26(31):8195-205.

    Monteiro P, Gilot D, Le Ferrec E, Rauch C, Lagadic-Gossmann D, Fardel O. Dioxin-mediated up-regulation of aryl hydrocarbon receptor target genes is dependent on the calcium/calmodulin/CaMKIalpha pathway. Mol Pharmacol. 2008 Mar;73(3):769-77. Epub 2007 Dec 18.

    Kolokoltsov AA, Deniger D, Fleming EH, Roberts NJ Jr, Karpilow JM, Davey RA. Small interfering RNA profiling reveals key role of clathrin-mediated endocytosis and early endosome formation for infection by respiratory syncytial virus. J Virol. 2007 Jul;81(14):7786-800. Epub 2007 May 9.

    Hussain KM, Leong KL, Ng MM, Chu JJ. The essential role of clathrin-mediated endocytosis in the infectious entry of human enterovirus 71. J Biol Chem. 2011 Jan 7;286(1):309-21. Epub 2010 Oct 18.

     

  2. General Screening References

    Parsons BD, Schindler A, Evans DH, Foley E. A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay.  PLoS One. 2009 Dec 29;4(12):e8471.

    Jiang M, Instrell R, Saunders B, Berven H, Howell M. Tales from an academic RNAi screening facility; FAQs. Brief Funct Genomics. 2011 Jul;10(4):227-37. doi: 10.1093/bfgp/elr016. Epub 2011 Apr 28.