RAPID GENERATION OF RAAV PACKAGING CELL LINES USING FACS

基因疗法已被提出用于治疗疾病 从癌症、病毒感染到代谢紊乱 例如囊性纤维化。 有效基因治疗的主要障碍 仍然是以治疗剂量输送基因药物。 腺- 相关的病毒载体可用作基因递送载体 应用程序的数量。 使用这种方法的主要限制是 技术正在实现可重复的载体制剂， 感染性滴度和低缺陷颗粒比率。 到 要想成功开发这项技术，整个过程必须 可规模化生产，产品一致，无污染 腺病毒和野生型AAV。 Immusol计划开发 使用流式细胞术生产rAAV载体的包装细胞系 流式细胞术以连续监测转基因表达。 帮手 用于rAAV载体生产的质粒，其并入了FACS可分选 将构建标记并用于检测高滴度载体 在瞬时测定中的生产。 稳定的包装细胞系， 这些质粒将被开发和筛选， 必要的辅助函数的表达式。 包装细胞系 具有所需滴度和颗粒比率的细胞将适于在 悬浮培养和在低血清或无血清培养基中培养。 建议的商业应用 Immusol计划商业化开发重组Adeno- 用于基因递送的相关病毒载体包装细胞系， 包括核酶递送，用于各种疾病， 传染病和代谢疾病以及癌症。

Gene therapy has been proposed in the treatment of diseases ranging from cancer and viral infections to metabolic disorders such as cystic fibrosis. The main hurdle to effective gene therapy remains delivery of genetic drugs at therapeutic doses. Adeno- associated viral vectors can be used as gene delivery vehicles in a number of applications. The major limitation to the use of this technology is achieving reproducible vector preparations with high infectious titer and a low ratio of defective particles. To successfully develop this technology, the entire process must be scaleable and yield consistent product free of contaminating adenovirus and wild-type AAV. Immusol plans to develop packaging cell lines for rAAV vector production using flow cytometry to continuously monitor transgene expression. Helper plasmids for rAAV vector production incorporating FACS sortable markers will be constructed and used to test for high titer vector production in transient assays. Stable packaging cell lines based on these plasmids will be developed and screened for stable expression of the necessary helper functions. Packaging cell lines with desirable titer and particle ratios will be adapted for growth in suspension culture and in low or no serum media. PROPOSED COMMERCIAL APPLICATION Immusol plans to commercially develop recombinant Adeno- associated viral vector packaging cell lines for use in gene delivery, including ribozyme delivery, for assorted diseases including infectious and metabolic disease and cancer.