4.8. Green Fluorescent Protein (GFP) Growth Competition Assay

To validate the results of the high-throughput screens, BL cell lines were infected by individual miRZip, pCDH, or CRISPR/Cas9 constructs, aiming at an infection efficiency of 20% to 50% GFP+ cells on day 4 or day 6. The percentage of GFP+ cells was monitored by flow cytometry (BD Biosciences, San Jose, CA, USA), for a period of 22 days. GFP+ percentages were normalized to the percentage of GFP+ cells at day 4 (miRZip) or day 6 (pCDH and CRISPR/Cas9). Statistical analysis of GFP competition assays was performed, as described previously [55]. In brief, decrease in percentages of GFP+ cells over time was compared with the controls, using a mixed model, with time and the interaction of time and construct types as fixed effects and the measurement repeat within construct types as random effect in SPSS (22.0.0.0 version, IBM, Armonk, New York, USA).