MAY 12, 2015 8:00 AM PDT

RNAi success in difficult-to-transfect cells by self-delivering siRNA

Sponsored by: Dharmacon, Dharmacon
Speakers

Abstract

Please click here to watch this webinar On Demand


siRNA-mediated silencing of gene expression has revolutionized the study of biology by enabling rapid and unbiased loss-of-function studies to be carried out in numerous cell types. However, siRNA application in difficult-to-transfect cell types, like primary or suspension cells, is very labor-intensive and technically challenging. Lentiviral transduction and electroporation offer transfection alternatives, but can result in unwanted cell death or viral responses.


Novel, chemically-modified siRNAs offer a solution to this problem, as these siRNAs enter into difficult-to-transfect cell types without the need of a delivery reagent and are available in many formats, including libraries for RNAi screening. In this webinar we will introduce Dharmacon Accell siRNA and present a study where we have screened an Accell siRNA library targeting the expression of 72 distinct genes in conjunction with a High Content Image Analysis platform as a proof-of-principle strategy to identify genes involved in LFA-1-mediated migration in primary human T cells. This study demonstrates the ease and benefits of conducting siRNA library screens in primary human T cells using self-delivering Accell siRNAs and emphasizes the feasibility and potential of this approach for elucidating the signaling pathways that regulate T cell function.


Webinar participants will learn about:



  • Availability of novel siRNA tools for gene silencing in difficult-to-transfect cells

  • Successful non-viral RNAi results in primary neurons, brain slices, and in vivo brain

  • Methods and results of a high-content siRNA screen in primary T-cells


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