MAY 12, 2015 08:00 AM PDT

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

SPONSORED BY: Dharmacon, Dharmacon
9 54 16337

  • Senior Product Manager, GE Healthcare
      The speaker is Louise Baskin, MS, senior product manager, GE Healthcare. Louise, who joined Dharmacon RNA Technologies in 2005, has led the development and commercial launch of multiple genome-wide siRNA and microRNA product lines. She is also responsible for developing and expanding the Dharmacon Edit-R Genome Engineering portfolio, which includes synthetic crRNA and lentiviral sgRNA, in addition to Cas9 nucleases in inducible or constitutive lentiviral vectors, DNA plasmid, mRNA, or protein formats. She works closely with a respected team of R&D scientists on innovative CRISPR-Cas9 tools and services. Louise earned a master's degree in molecular biology and genetics at Northwestern University.
    • Senior Research Fellow in the Dept. of Clinical Medicine,Trinity College
        Michael Freeley is a Senior Research Fellow in the Dept. of Clinical Medicine in Trinity College Dublin, Ireland. His research focuses on the cell biology of the T cell immune response, particularly the signalling pathways and proteins that regulate T cell activation and migration. He utilises a number of research tools such as siRNAs, screening of RNAi libraries and High Content Analysis to address many of these questions. Characterisation of the pathways that regulate T cell activation and migration holds great promise for modulating T cell immune responses in inflammatory/autoimmune diseases, infection and cancer. Michael's research findings have been published in high-impact peer-reviewed international journals, including The Journal of Immunology, The Journal of Biomolecular Screening, Biochemical Journal, Science, The Journal of Biological Chemistry and Journal of Immunological Methods. Michael and colleagues were the recipients of the ‘High Content Analysis' award in 2010 (sponsored by General Electric) for their work on a High Content multi-parametric analysis of T cell migration. Michael holds a PhD in Biochemistry from the National University of Ireland (Royal College of Surgeons in Ireland, 2004) and a first-class honours degree in Biotechnology from Dublin City University (1999).


      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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