FEB 20, 2017 08:00 AM PST

Presenting a complete workflow - from start to finish!

Speakers
  • Senior Scientist, Biology Research and Development, Dharmacon
    Biography
      Emily Anderson is a Senior Scientist working in gene modulation research at Dharmacon (part of GE Healthcare) located in Lafayette, Colorado. She received her B.S. in Chemistry from the University of Rochester and her Ph.D. in Biochemistry from the University of Colorado at Boulder, where she elucidated the structure and function of protein:single-stranded DNA interactions at telomeres. In 2003 Emily joined Dharmacon and has worked extensively on the mechanism, function, specificity, and delivery of RNA interference (RNAi) and its applications. She is an inventor on several issued patents, has contributed to the introduction of multiple product lines, and has authored peer-reviewed publications in the RNAi field. In her current role, she manages several projects involving the development and application of gene engineering and other gene modulation technologies.

    Abstract:

    The CRISPR-Cas9 system is being widely used for genome engineering in many different biological applications. It was originally adapted from the bacterial Type II CRISPR system and uses a Cas9 endonuclease guided by RNA to introduce double-strand DNA breaks at specific locations in the genome. The Dharmacon Edit-R CRISPR-Cas9 Gene Engineering platform includes the three components required for gene editing in mammalian cells: (1) a plasmid expressing a Cas9 nuclease, (2) a chemically synthesized tracrRNA, and (3) a synthetic crRNA designed to the target site of interest. In this presentation, the complete workflow of the Edit-R platform is demonstrated to knock out the PPIB gene at the protein level in HEK293T cells. Data from the analysis of 42 edited clones are presented.
      
    Attendees will learn:

    • CRISPR-Cas9 background and crRNA design considerations
    • Transfection optimization for maximal editing efficiency
    • Enrichment of cell populations with gene editing events by FACS
    • Clonal selection
    • Detailed analysis of editing events

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