de Fiebre C M, Wu P, Notabartolo D, Millard W J, Meyer E M
Department of Pharmacology and Therapeutics, University of Florida College of Medicine, J. Hillis Miller Health Center, Gainesville 32610-0267.
Neurochem Res. 1994 Jun;19(6):643-8. doi: 10.1007/BF00967701.
The ability of Sendai virosomes or Lipofectin to introduce an AAV vector into primary rat brain astroglial cultures was characterized. The pJDT95npy vector was constructed by inserting rat NPY cDNA downstream from the indigenous AAV p5, p19 and p40 promoters in pJDT95. Lipofectin-mediated transfection with pJDT95npy (10 micrograms) resulted in pronounced expression of several NPY mRNA species: p5-driven (3.3 kb), p19-driven (2.7 kb) and p40-driven (0.6, 0.8, 1.1, and 1.8 kb). Exposure to virosomally encapsulated pJDT95npy (50 or 100 ng) resulted in transient expression of some p40-driven mRNA species (0.8 and 1.8 kb). Neither method produced astroglia cells which synthesized mature NPY immunoreactivity. This demonstrates that an AAV-derived vector can drive gene expression in astroglia, that Sendai virosomes can infuse vectors into astroglia, but that the amount of DNA infused in this manner may limit long term expression.
对仙台病毒体或脂质体将腺相关病毒(AAV)载体导入原代大鼠脑星形胶质细胞培养物中的能力进行了表征。pJDT95npy载体是通过将大鼠神经肽Y(NPY)cDNA插入pJDT95中天然AAV的p5、p19和p40启动子下游构建而成。用pJDT95npy(10微克)进行脂质体介导的转染导致几种NPY mRNA种类的明显表达:p5驱动的(3.3 kb)、p19驱动的(2.7 kb)和p40驱动的(0.6、0.8、1.1和1.8 kb)。暴露于病毒体包裹的pJDT95npy(50或100纳克)导致一些p40驱动的mRNA种类(0.8和1.8 kb)的瞬时表达。两种方法均未产生合成成熟NPY免疫反应性的星形胶质细胞。这表明源自AAV的载体可驱动星形胶质细胞中的基因表达,仙台病毒体可将载体注入星形胶质细胞,但以这种方式注入的DNA量可能会限制长期表达。