Efficient transduction of human neurons with an adeno-associated virus vector.
Efficient transduction of human neurons with an adeno-associated virus vector.
复制标题
用腺相关病毒载体有效转导人类神经元。
作者：
B. Du;P. Wu;D. Boldt‐Houle;E. Terwilliger
腺相关病毒载体含有由CMV立即早期启动子（AAV β-gal）驱动的lacZ基因进行了评价，其转导效率和整合能力在nondividing人NT神经元。免疫细胞化学证实了AAV β-gal转导效率的剂量依赖性模式，高达100%的神经元表达基因产物。未检测到载体的神经毒性作用。来自转导的神经元的高分子量细胞DNA的定量PCR分析表明，AAV β-gal基因组的拷贝数以时间依赖性方式逐渐增加，表明在转导后约1周的时间内载体整合的缓慢但渐进的速率。与进入标准靶细胞系相比，AAV β-gal进入NT神经元的转导效率相等或更高，表明NT神经元容易对AAV介导的基因转移敏感。这项研究表明，基于AAV的载体可以有效地在有丝分裂后的人类神经元中稳定表达外源基因。
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.