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.
Highlights
The ongoing development of tools for targeted genome editing using CRISPR has revolutionized functional genomics (Shalem et al, 2015)
CRISPR-based forward genetic screens in pooled format have largely been used to study gene essentiality and synthetic lethality in different contexts (Shalem et al, 2014; Hart et al, 2015; Wang et al, 2015, 2017). These screens are largely restricted to measuring growth phenotypes or to phenotype/cellular markers that can be selected by fluorescence-activated cell sorting (FACS) and require next-generation sequencing (NGS) based readouts to analyze the data
In solid-phase transfection, the microwell plates are coated with the transfection reagent and the synthetic crRNA:tracrRNA complexes that are stabilized by sucrose and gelatin
Summary
Özdemirhan Serçin , Sabine Reither, Paris Roidos , Nadja Ballin , Spyridon Palikyras, Anna Baginska, Katrin Rein, Maria Llamazares , Aliaksandr Halavatyi, Hauke Winter, Thomas Muley, Renata Z Jurkowska , Amir Abdollahi6,7 , Frank T Zenke, Beate Neumann2 & Balca R Mardin1,*
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