Authors
Benjamin Steyer, Jared Carlson-Stevermer, Nicolas Angenent-Mari, Andrew Khalil, Ty Harkness, Krishanu Saha
Description
Non-viral gene-editing of human cells using the CRISPR-Cas9 system requires optimized delivery of multiple components. Both the Cas9 endonuclease and a single guide RNA, that defines the genomic target, need to be present and co-localized within the nucleus for efficient gene-editing to occur. This work describes a new high-throughput screening platform for the optimization of CRISPR-Cas9 delivery strategies. By exploitinghigh content image analysisand microcontactprintedplates, multi-parametricgene-editing outcome datafrom hundreds to thousandsof isolated cell populations can be screened simultaneously. Employing this platform, we systematically screened four commercially availablecationic lipid transfection materials with a range of RNAs encoding the CRISPR-Cas9 system. Analysis of Cas9 expression and editing of a fluorescent mCherry reporter transgene within human embryonic kidney cells was
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