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APR 05, 2017 6:00 AM PDT

A workflow for CRISPR-Cas9 high throughput arrayed screening with synthetic crRNA

Speaker
  • Senior Product Manager, GE Healthcare
    Biography
      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.

    Abstract

    There is rapidly growing interest in using the CRISPR-Cas9 system for functional screening, both as a primary screening tool and as an orthogonal tool for RNAi hit validation. High throughput synthesis, combined with a proprietary algorithm that selects highly functional and specific guide RNAs, allows rapid generation of CRISPR RNA (crRNA) collections in arrayed formats. Arrayed crRNA screens offer the advantage of applying sophisticated assays, such as those requiring high-content microscopy, to investigate phenotypes that involve intracellular localization, morphological changes, or require time-lapse investigation. While pooled lentiviral sgRNA screens have demonstrated utility for loss-of-function studies, the types of assays used are typically limited to viability, sensitization, and some reporter assays. We will demonstrate the application of arrayed screening with synthetic crRNA libraries across multiple assay types, and present considerations for experimental success.


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