A solid-phase transfection platform for arrayed CRISPR screens.
Özdemirhan SerçinSabine ReitherParis RoidosNadja BallinSpyridon PalikyrasAnna BaginskaKatrin ReinMaria LlamazaresAliaksandr HalavatyiHauke WinterThomas MuleyRenata Z JurkowskaAmir AbdollahiFrank T ZenkeBeate NeumannBalca R MardinPublished in: Molecular systems biology (2020)
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.
Keyphrases
- high throughput
- genome wide
- crispr cas
- genome editing
- dna methylation
- copy number
- single cell
- endothelial cells
- induced apoptosis
- papillary thyroid
- gene expression
- randomized controlled trial
- oxidative stress
- squamous cell carcinoma
- signaling pathway
- clinical trial
- cell cycle arrest
- lymph node metastasis
- loop mediated isothermal amplification