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Lipid mediated gene delivery in the adult rat brain: quantitative analysis of expression.

机译:成年大鼠脑中脂质介导的基因传递:表达的定量分析。

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摘要

Gene transfer into the adult brain is potentially an attractive alternative to commonly employed transgenic approaches. DNA-lipid complexes have been used to obtain brain gene transfer, but data are sparse to indicate to what extent this results in significant expression of functional protein. Here, an expression construct encoding the functional reporter, chloramphenicol-acetyl-transferase (CAT), was complexed to a novel biodegradable lipid, and delivered into the rat brain. CAT-activity was assayed in tissue extracts to allow a precise quantitation of functional enzyme protein. Following bilateral intraventricular (i.c.v.) injection, robust enzyme activity was found in all brain regions studied, peaking at 4 weeks. Other routes of administration, e.g. intra-parenchymal injection or chronic infusion of complexes, resulted in marginal or no activity. Presence of CAT mRNA and plasmid DNA in tissue extracts was confirmed at 4 weeks post i.c.v. administration. In agreement with previous studies, labelled lipid-DNA complexes were mainly found in the ventricular ependyma. Present data support the feasibility of lipid mediated brain gene transfer, and outline some of its anatomical and temporal limitations.
机译:基因转移到成人大脑中可能是常用转基因方法的一种有吸引力的替代方法。 DNA-脂质复合物已用于获得脑基因转移,但数据稀少,表明这会在多大程度上导致功能蛋白的显着表达。在此,将编码功能性报告基因氯霉素-乙酰基转移酶(CAT)的表达构建体复合到新型可生物降解的脂质上,并递送至大鼠脑中。在组织提取物中检测CAT活性,以精确定量功能性酶蛋白。在双侧脑室内(i.c.v.)注射后,在所有研究的大脑区域均发现了强大的酶活性,在4周时达到峰值。其他给药途径,例如实质内注射或慢性输注复合物导致边缘活动或无活动。静脉注射后4周证实组织提取物中CAT mRNA和质粒DNA的存在。行政。与先前的研究一致,标记的脂质-DNA复合物主要存在于心室室管膜。目前的数据支持脂质介导的脑基因转移的可行性,并概述了其在解剖学和时间上的局限性。

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