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A solid-phase transfection platform for arrayed CRISPR screens.

ABSTRACT: Arrayed CRISPR-based screens emerge as a powerful alternative to pooled screens making it possible to investigate a wide range of cellular phenotypes that are typically not amenable to pooled screens. Here, we describe a solid-phase transfection platform that enables CRISPR-based genetic screens in arrayed format with flexible readouts. We demonstrate efficient gene knockout upon delivery of guide RNAs and Cas9/guide RNA ribonucleoprotein complexes into untransformed and cancer cell lines. In addition, we provide evidence that our platform can be easily adapted to high-throughput screens and we use this approach to study oncogene addiction in tumor cells. Finally demonstrating that the human primary cells can also be edited using this method, we pave the way for rapid testing of potential targeted therapies.

SUBMITTER: Sercin O

PROVIDER: S-EPMC6926425 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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A solid-phase transfection platform for arrayed CRISPR screens.

Serçin Özdemirhan Ö Reither Sabine S Roidos Paris P Ballin Nadja N Palikyras Spyridon S Baginska Anna A Rein Katrin K Llamazares Maria M Halavatyi Aliaksandr A Winter Hauke H Muley Thomas T Jurkowska Renata Z RZ Abdollahi Amir A Zenke Frank T FT Neumann Beate B Mardin Balca R BR

Molecular systems biology 20191201 12

Arrayed CRISPR-based screens emerge as a powerful alternative to pooled screens making it possible to investigate a wide range of cellular phenotypes that are typically not amenable to pooled screens. Here, we describe a solid-phase transfection platform that enables CRISPR-based genetic screens in arrayed format with flexible readouts. We demonstrate efficient gene knockout upon delivery of guide RNAs and Cas9/guide RNA ribonucleoprotein complexes into untransformed and cancer cell lines. In ad ...[more]

PMID: 31885201

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