Figure 8
From: Engineered CHO cells as a novel AAV production platform for gene therapy delivery
Development and selection of HSV-1 producer CHO-ICP27 pools and clones. (a) Construct for development of stable CHO-HV1-ICP27 pools using random integration. The Chinese hamster codon-optimized HSV-1 ICP27 ORF expression was driven by CMV promoter under puromycin selection. (b) Construct for development of stable CHO-HV1-ICP27 pools using CRISPR/Cas9 technology. Donor plasmid was constructed and encoded codon-optimized Chinese hamster ICP27 and puromycin cassettes flanked by two homology arms. (c) Mean fluorescence intensity (MFI) of ICP27 expression from the final selected clones. Data (n = 2 per sample) are expressed as mean ± SD. (d) Comparison of rHSV-AAV9 production in CHO-HV1-ICP27-C11 and V27 cells using different MOIs. Data were analyzed with two-way ANOVA and reported as mean ± SD.
