A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: Polyethylenimine
A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: Polyethylenimine
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DOI：
10.1089/hum.1996.7.16-1947
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发表时间：
1996-10-20
影响因子：
4.2
通讯作者：
Demeneix, BA
中科院分区：
文献类型：
--
作者：
Abdallah, B;Hassan, A;Demeneix, BA
非病毒基因转移到中枢神经系统（CNS）为神经系统疾病提供了安全的治疗方法和操纵基因表达以研究神经功能提供了前景。然而，迄今为止报道的结果令人失望。我们发现阳离子聚合物聚乙烯亚胺（PEI）在成熟小鼠脑中提供了前所未有的高水平的转基因表达。用25-kD聚合物获得了最高水平的转染。用这种制备物，具有最接近中性的平均离子电荷比的DNA/PEI复合物给出了最好的结果。在这种条件下，并使用巨细胞病毒（CMV）-荧光素酶构建物，我们获得了高达0.4 × 10（6）RLU/μ g DNA（相当于0.4 ng荧光素酶），这接近使用PEI转染神经元培养物和更容易转染的新生小鼠大脑获得的值（10（6）RLU/μ g DNA），在注射到大脑皮层、海马和下丘脑后，标记物（荧光素酶）或功能基因（belt）在神经元和神经胶质细胞中得到广泛表达（超过6 mm（3））。注射后3个月以上，在皮层神经元中发现转基因表达，使用任何制剂均未观察到发病率，因此，PEI，一种低毒性载体，似乎具有用于脑的基础研究和基因治疗的潜力。
Nonviral gene transfer into the central nervous system (CNS) offers the prospect of providing safe therapies for neurological disorders and manipulating gene expression for studying neuronal function, However, results reported so far have been disappointing, We show that the cationic polymer polyethylenimine (PEI) provides unprecedentedly high levels of transgene expression in the mature mouse brain, Three different preparations of PEI (25-, 50-, and 800-kD) were compared for their transfection efficiencies in the brains of adult mice, The highest levels of transfection were obtained with the 25-kD polymer, With this preparation, DNA/PEI complexes bearing mean ionic charge ratios closest to neutrality gave the best results, Under such conditions, and using a cytomegalovirus (CMV)-luciferase construction, we obtained up to 0.4 10(6) RLU/mu g DNA (equivalent to 0.4 ng of luciferase), which is close to the values obtained using PEI to transfect neuronal cultures and the more easily transfected newborn mouse brain (10(6) RLU/mu g DNA), Widespread expression (over 6 mm(3)) of marker (luciferase) or functional genes (belt) was obtained in neurons and glia after injection into the cerebral cortex, hippocampus, and hypothalamus. Transgene expression was found more than 3 months post-injection in cortical neurons, No morbidity was observed with any of the preparations used, Thus, PEI, a low-toxicity vector, appears to have potential for fundamental research and genetic therapy of the brain.