An image

An image

Imagine performing an RNA interference (RNAi) screen and identifying no novel hits. It can happen1 
 
Failure to uncover new hits, and a high false positive hit rate are common in large RNAi screens
2,3. These stem from wide-spread off-target effects of siRNAs. Off-targeting not only incurs a significant drain on time and resources from validation processes required to weed out false hits, it also increases assay noise which limits the detection of true hits.

siPOOLs efficiently counter off-targets and increase reliability of gene silencing. RNAi screening with the siPOOL human kinase library: 
  • reduces false positives
  • improves detection of true hits
  • improves consistency between screens
  • avoids inefficient testing of multiple siRNAs
  • eases data analysis
  • obtains results quickly
  • can be applied to many cell lines
  • can be multiplexed with other treatments/siPOOLs to uncover genetic interactions (e.g. synthetic lethality screens)
 

Avoid false positives, increase true hits

An image
Transcriptome-wide expression after siPOOL or siRNA treatment (HeLa cells, 3 nM, 48 h)

Just use once for reliable phenotypes

An image
Cell viability correlation between sequence-independent siRNAs/siPOOLs targeting the same gene (36 genes, A549 cells, 3 nM, 72 h)

References

1 Sigoillot, F. D., Lyman, S., Huckins, J. F., Adamson, B., Chung, E., Quattrochi, B., and King, R. W. (2012) A bioinformatics method identifies prominent off-targeted transcripts in RNAi screens. Nat. Methods. 9, 363–366

Bushman, F. D., Malani, N., Fernandes, J., D’Orso, I., Cagney, G., Diamond, T. L., Zhou, H., Hazuda, D. J., Espeseth, A. S., König, R., Bandyopadhyay, S., Ideker, T., Goff, S. P., Krogan, N. J., Frankel, A. D., Young, J. A. T., and Chanda, S. K. (2009) Host Cell Factors in HIV Replication: Meta-Analysis of Genome-Wide Studies. PLoS Pathog. 5, e1000437

Hasson, S. A., Kane, L. A., Yamano, K., Huang, C.-H., Sliter, D. A., Buehler, E., Wang, C., Heman-Ackah, S. M., Hessa, T., Guha, R., Martin, S. E., and Youle, R. J. (2013) High-content genome-wide RNAi screens identify regulators of parkin upstream of mitophagy. Nature. 504, 291–295

4 Welsbie, D. S., Mitchell, K. L., Jaskula-Ranga, V., Sluch, V. M., Yang, Z., Kim, J., Buehler, E., Patel, A., Martin, S. E., Zhang, P. W., Ge, Y., Duan, Y., Fuller, J., Kim, B. J., Hamed, E., Chamling, X., Lei, L., Fraser, I. D. C., Ronai, Z. A., Berlinicke, C. A., and Zack, D. J. (2017) Enhanced Functional Genomic Screening Identifies Novel Mediators of Dual Leucine Zipper Kinase-Dependent Injury Signaling in Neurons. Neuron. 94, 1142–1154.e6

LATEST NEWS! siPOOLs in the human kinase library are also available for individual purchase at the 2 nmol scale.
 


If you would like to receive more information, request a quotation or have your questions addressed by a scientist, please fill in all relevant fields:
    


We provide:
  • 505 siPOOLs against classical human protein kinases, custom extensions possible
  • Provided in 96/384-well plates or 2D barcoded tubes. Lyophilized or in solution. Scales: 0.1-5 nmol 
  • Plate arraying service and additional controls
  • Data analysis/bioinformatics service
  • Rescue constructs for secondary assays
  • Affordable siPOOL re-orders
 
 

User Review




Dr. Derek S Welsbie 
Assistant Professor  
University of California, San Diego, USA

“Our lab uses arrayed, high-throughput functional genomic screening in primary neurons to identify potential neuroprotective drug targets. Having tested over 75,000 siRNA sequences, it is quite apparent that off-target effects dominate siRNA-mediated phenotypes. In contrast, in our hands, siPOOLs have much greater predictive power in that phenotypes we see with these can be reproduced using cells containing conventional knockouts for the same genes4. We now routinely use siPOOLs and are moving away from single siRNAs.”


siPOOL human kinase library brochure