A solid‐phase transfection platform for arrayed CRISPR screens

• Published in: Molecular systems biology 2019-12, Vol.15 (12), p.e8983-n/a
• Main Authors: Serçin, Özdemirhan, Reither, Sabine, Roidos, Paris, Ballin, Nadja, Palikyras, Spyridon, Baginska, Anna, Rein, Katrin, Llamazares, Maria, Halavatyi, Aliaksandr, Winter, Hauke, Muley, Thomas, Jurkowska, Renata Z, Abdollahi, Amir, Zenke, Frank T, Neumann, Beate, Mardin, Balca R
• Format: Article
• Language: English
• Subjects: Addictions; arrayed screens; Cell adhesion & migration; Cell cycle; Cell Line, Tumor; CRISPR; CRISPR-Cas Systems; CRISPR/Cas9; Cytotoxicity; Deoxyribonucleic acid; DNA; EMBO09; EMBO22; Experiments; Flow cytometry; Gene Editing - instrumentation; Genes; Genetic Predisposition to Disease; Genetic screening; Genomics; gRNA/RNP delivery; High-Throughput Screening Assays; Humans; Kinases; Method; Methods; Microscopy; Morphology; Neoplasms - genetics; Phenotype; Phenotypes; Proteins; Ribonucleic acid; RNA; RNA, Guide, CRISPR-Cas Systems - pharmacology; solid phase; Transfection; transfection platform; Tumor cell lines; Tumor cells
• ISSN: 1744-4292; 1744-4292
• DOI: 10.15252/msb.20198983

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. Synopsis This study presents a solid phase transfection platform that enables CRISPR‐based genetic screens in arrayed format with flexible readouts. Solid phase transfection of gRNAs or RNPs is applicable to a wide variety of cell lines. Solid phase transfection can be easily adapted to high throughput screens. A demonstration that human primary cells can be edited using this platform, indicates the potential of the approach for rapid testing of candidate targeted therapies. Graphical Abstract This study presents a solid phase transfection platform that enables CRISPR‐based genetic screens in arrayed format with flexible readouts.