一种提高单链腺相关病毒载体在体外和体内转导效率的简单方法。
A simple method to increase the transduction efficiency of single-stranded adeno-associated virus vectors in vitro and in vivo.
机构信息
Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32610, USA.
出版信息
Hum Gene Ther. 2011 May;22(5):633-40. doi: 10.1089/hum.2010.243.
Abstract
We have recently shown that co-administration of conventional single-stranded adeno-associated virus 2 (ssAAV2) vectors with self-complementary (sc) AAV2-protein phosphatase 5 (PP5) vectors leads to a significant increase in the transduction efficiency of ssAAV2 vectors in human cells in vitro as well as in murine hepatocytes in vivo. In the present study, this strategy has been further optimized by generating a mixed population of ssAAV2-EGFP and scAAV2-PP5 vectors at a 10:1 ratio to achieve enhanced green fluorescent protein (EGFP) transgene expression at approximately 5- to 10-fold higher efficiency, both in vitro and in vivo. This simple coproduction method should be adaptable to any ssAAV serotype vector containing transgene cassettes that are too large to be encapsidated in scAAV vectors.
摘要
我们最近发现，将传统的单链腺相关病毒 2（ssAAV2）载体与自我互补（sc）AAV2-蛋白磷酸酶 5（PP5）载体联合使用，可显著提高 ssAAV2 载体在体外人细胞以及体内小鼠肝细胞中的转导效率。在本研究中，通过生成 10:1 比例的 ssAAV2-EGFP 和 scAAV2-PP5 载体的混合群体，进一步优化了该策略，以实现体外和体内增强型绿色荧光蛋白（EGFP）转基因表达效率提高约 5-10 倍。这种简单的共生产方法应该适用于任何含有太大而无法封装在 scAAV 载体中的转基因盒的 ssAAV 血清型载体。
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