Orlans Harry O, Edwards Thomas L, De Silva Samantha R, Patrício Maria I, MacLaren Robert E
Nuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford, UK.
Oxford Eye Hospital, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.
Methods Mol Biol. 2018;1715:289-303. doi: 10.1007/978-1-4939-7522-8_21.
Recombinant adeno-associated viral (AAV) vectors have been successfully employed as the mode of gene delivery in several clinical trials for the treatment of inherited retinal diseases to date. The design of such vectors is critical in determining cellular tropism and level of subsequent gene expression that may be achieved following viral delivery. Here we describe a system for living retinal tissue extraction, ex vivo culture, viral transduction and assessment of transgene expression that may be used to assess viral constructs for gene therapy in the human retina at a preclinical stage.
重组腺相关病毒（AAV）载体已成功用作基因递送方式，用于迄今为止的多项治疗遗传性视网膜疾病的临床试验。此类载体的设计对于确定细胞嗜性以及病毒递送后可能实现的后续基因表达水平至关重要。在此，我们描述了一种用于活体视网膜组织提取、体外培养、病毒转导和转基因表达评估的系统，该系统可用于在临床前阶段评估用于人类视网膜基因治疗的病毒构建体。
