Li Ying, Yang Yi, Wang Shu
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos #04-01, Singapore 138669.
Exp Physiol. 2005 Jan;90(1):39-44. doi: 10.1113/expphysiol.2004.028217. Epub 2004 Nov 12.
Recombinant baculoviruses have been employed as gene delivery vectors for mammalian cells, including neurones, during recent years. The aim of the current study was to develop a new recombinant baculovirus vector capable of enhancing gene expression in neurones. A hybrid promoter constructed by fusing the enhancer of human cytomegalovirus (CMV) immediately early promoter to the human platelet-derived growth factor (PDGF) beta-chain promoter was placed into a baculovirus expression cassette. In cultured neurones, baculovirus vectors containing the hybrid promoter augmented transgene expression up to 100-fold greater than that mediated by titre-matched baculovirus vectors with the PDGF promoter alone. Double immunostaining of tissue sections collected from the striatum and the retina injected with the new baculovirus vector demonstrated its specificity in driving gene expression almost exclusively in neurones, confirming the feasibility of using a tissue-specific promoter in the context of baculovirus vectors to provide cell type-specific transgene expression. The attributes of the new baculovirus vector might have practical implications for gene therapy in the nervous system.
近年来,重组杆状病毒已被用作包括神经元在内的哺乳动物细胞的基因传递载体。本研究的目的是开发一种能够增强神经元基因表达的新型重组杆状病毒载体。通过将人巨细胞病毒(CMV)立即早期启动子的增强子与人血小板衍生生长因子(PDGF)β链启动子融合构建的杂合启动子被置于杆状病毒表达盒中。在培养的神经元中,含有杂合启动子的杆状病毒载体增强转基因表达的能力比仅带有PDGF启动子且滴度匹配的杆状病毒载体介导的表达高出100倍。对注射了新型杆状病毒载体的纹状体和视网膜组织切片进行双重免疫染色,结果表明其在驱动基因表达方面具有特异性,几乎仅在神经元中表达,这证实了在杆状病毒载体中使用组织特异性启动子以提供细胞类型特异性转基因表达的可行性。新型杆状病毒载体的特性可能对神经系统的基因治疗具有实际意义。