Du B, Wu P, Boldt-Houle D M, Terwilliger E F
Division of Infectious Disease, New England Deaconess Hospital, Harvard Medical School, Boston, MA, USA.
Gene Ther. 1996 Mar;3(3):254-61.
An adeno-associated virus vector containing a lacZ gene driven by a CMV immediate-early promoter (AAV beta-gal) was evaluated with respect to its transduction efficiency and integration ability in nondividing human NT neurons. A dose-dependent pattern in transduction efficiency of the AAV beta-gal was demonstrated immunocytochemically, with up to 100% of the neurons expressing the gene product. No neurotoxic effects of the vector were detected. Quantitative PCR analyses of high molecular weight cellular DNA from the transduced neurons indicated that the copy number of the AAV beta-gal genome increased gradually in a time dependent manner, suggesting a slow but progressive rate of vector integration over a period of approximately 1 week following transduction. Equal or greater transduction efficiency of the AAV beta-gal into NT neurons than into a standard target cell line indicated that the NT neurons were readily susceptible to AAV-mediated gene transfer. This study demonstrates that AAV-based vectors can efficiently transduce and stably express a foreign gene in post-mitotic human neurons.
对一种由巨细胞病毒立即早期启动子驱动的含lacZ基因的腺相关病毒载体（腺相关病毒β-半乳糖苷酶）在非分裂的人NT神经元中的转导效率和整合能力进行了评估。通过免疫细胞化学方法证明了腺相关病毒β-半乳糖苷酶转导效率呈剂量依赖性模式，高达100%的神经元表达该基因产物。未检测到该载体的神经毒性作用。对转导神经元的高分子量细胞DNA进行的定量PCR分析表明，腺相关病毒β-半乳糖苷酶基因组的拷贝数随时间呈逐渐增加的趋势，这表明在转导后的大约1周时间内载体整合速度缓慢但呈渐进性。腺相关病毒β-半乳糖苷酶对NT神经元的转导效率等于或高于对标准靶细胞系的转导效率，这表明NT神经元很容易受到腺相关病毒介导的基因转移的影响。这项研究表明，基于腺相关病毒的载体能够在有丝分裂后的人神经元中高效转导并稳定表达外源基因。
