合成生物学
生物制造
生物
转染
衣壳
细胞培养
质粒
遗传增强
计算生物学
基因组
HEK 293细胞
基因
病毒
辅助病毒
病毒学
基因传递
遗传学
作者
Zion Lee,Min Lü,Eesha Irfanullah,Morgan Soukup,Wei‐Shou Hu
标识
DOI:10.1021/acssynbio.2c00207
摘要
Recombinant adeno-associated viruses (rAAV) are important gene delivery vehicles for gene therapy applications. Their production relies on plasmid transfection or virus infection of producer cells, which pose a challenge in process scale-up. Here, we describe a template for a transfection-free, helper virus-free rAAV producer cell line using a synthetic biology approach. Three modules were integrated into HEK293 cells including an rAAV genome and multiple inducible promoters controlling the expression of AAV Rep, Cap, and helper coding sequences. The synthetic cell line generated infectious rAAV vectors upon induction. Independent control over replication and packaging activities allowed for manipulation of the fraction of capsid particles containing viral genomes, affirming the feasibility of tuning gene expression profiles in a synthetic cell line for enhancing the quality of the viral vector produced. The synthetic biology approach for rAAV production presented in this study can be exploited for scalable biomanufacturing.
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