Non-viral neuronal gene delivery mediated by the HC fragment of tetanus toxin
Non-viral neuronal gene delivery mediated by the HC fragment of tetanus toxin
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DOI：
10.1046/j.1432-1327.1999.00108.x
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发表时间：
1999-02-01
期刊：
影响因子：
--
通讯作者：
Fairweather, N
中科院分区：
文献类型：
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作者：
Knight, A;Carvajal, J;Fairweather, N
许多遗传性神经系统疾病和癌症可能受益于向中枢神经系统神经元的有效靶向基因递送。破伤风毒素的无毒片段C (H-C)保留了破伤风全毒素的特异性神经细胞结合和转运特性。 H-C 片段此前已被用于促进体外和体内神经元细胞摄取附着蛋白，例如辣根过氧化物酶、β-半乳糖苷酶和超氧化物歧化酶。我们报告了使用在酵母中产生的纯化重组 H-C 片段并与聚赖氨酸 [poly(K)] 共价结合以实现 DNA 的结合。我们证明，当用于转染细胞时，该构建体会在 N18 RE 105 细胞（神经母细胞瘤 x 神经胶质瘤小鼠/大鼠杂交细胞系）和 F98（神经胶质瘤细胞系）中产生非病毒基因递送和标记基因表达。转染依赖于 H-C 并且具有神经元细胞类型特异性。 H-C 可能被证明是未来神经元基因治疗的有用靶向配体。
Many inherited neurological diseases and cancers could potentially benefit from efficient targeted gene delivery to neurons of the central nervous system. The nontoxic fragment C (H-C) of tetanus toxin retains the specific nerve cell binding and transport properties of tetanus holotoxin. The H-C fragment has previously been used to promote the uptake of attached proteins such as horseradish peroxidase, beta-galactosidase and superoxide dismutase into neuronal cells in vitro and in vivo. We report the use of purified recombinant H-C fragment produced in yeast and covalently bound to polylysine [poly(K)] to enable binding of DNA. We demonstrate that when used to transfect cells, this construct results in nonviral gene delivery and marker gene expression in vitro in N18 RE 105 cells (a neuroblastoma x glioma mouse/rat hybrid cell line) and F98 (a glioma cell line). Transfection was dependent on H-C and was neuronal cell type specific. H-C may prove a useful targeting ligand for future neuronal gene therapy.