Automated microraft platform to identify and collect non-adherent cells successfully gene-edited with CRISPR-Cas9

Generation of cell lines with specific mutations is integral to the in vitro study of many diseases and the associated pathogenesis, and the CRISPR-Cas9 gene editing system has revolutionized the ability to efficiently generate disease models. Limiting dilution and FACS have traditionally been used to obtain clonal cell lines with specific genomic modifications introduced by the CRISPR-Cas9 system; however, they require large sample sizes and often yield low cell viability. The authors used the CellRaft® Technology to sort cells based on the temporal evolution of fluorescent protein expression (EGFP) to generate a CRISPR gene-edited cell line with a leukemia-associated mutation (S34F) in the U2AF1 protein, allowing for the further study of the consequences of this mutation on mRNA splicing in AML.