Flow cytometry-based PTM CRISPR-Cas9 screening of Rab22a-NeoF1 protein stability. Homo sapiens

1. Flow cytometry-based PTM CRISPR-Cas9 screening of Rab22a-NeoF1 protein stabilityU2OS pAd-DsRed-IRES-EGFP-Rab22a-NeoF1 cells infected lentivirus of Post translation modification CRISPR-Cas9 knock out library were sorted into GFP high and GFP low populations using FACS. Genomic DNA (gDNA) was isolated from GFP high and GFP low population cells, were subjected to the sgRNA PCR amplification and CRISPR screen data analysis.2. Flow cytometry-based kinome CRISPR-Cas9 screening of Rab22a-NeoF1 protein stabilityU2OS pAd-DsRed-IRES-EGFP-Rab22a-NeoF1 cells infected lentivirus of kinome CRISPR-Cas9 knock out library were sorted into GFP high and GFP low populations using FACS. Genomic DNA (gDNA) was isolated from GFP high and GFP low population cells, were subjected to the sgRNA PCR amplification and CRISPR screen data analysis.