Gene transfection by cationic liposome with a cationic cholesterol derivative

阳离子脂质体与阳离子胆固醇衍生物的基因转染

• 批准号: 12557207
• 负责人: NAKANISHI Mamoru
• 金额: $ 5.7万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557207/
• 关键词: Gene transfection; Cationic cholesterol; Cationic liposome; Non-viral vector; Microscopy; Endocytosis; Biosurfactant; Gene therapy; 非ウイルスベクター; 膜融合

We will demonstrate interesting cationic liposomes with novel cationic cholesterol derivatives, a new strategy in gene transfection developed by our group and the presently accepted molecular mechanism of gene transfection. Use of confocal laser scanning microscopy and atomic force microscopy in elucidating the molecular mechanism of gene transfection by cationic liposomes is also showed using examples from our own work. As delineated, both the confocal laser scanning microscopic and the atomic force microscopic results advocate for the involvement of the sequential three steps in gene transfection mediated by the cationic liposomes: endocytotic internalization of the lipoplexes into the target cells, endosome-lysosome fusion whereby the DNA gets released from the liposomes and moves towards the nucleus of the target cells and microtubule organization apparently involved in trafficking the transfected foreign genes to lysosomes. Furthermore, our experiment also shows couple of important strategies in gene transfection namely, use of liposomes made from biosurfactans and harnessing efficient gene transfection by activating the membrane-bound receptor molecules.

我们将展示具有新型阳离子胆固醇衍生物的有趣的阳离子脂质体，这是我们团队开发的一种新的基因转移策略，以及目前公认的基因转移的分子机制。利用共聚焦激光扫描显微镜和原子力显微镜阐明了阳离子脂质体基因转导的分子机制，并用我们自己的工作实例加以说明。如图所示，激光共聚焦扫描显微镜和原子力显微镜的结果都主张阳离子脂质体介导的基因转染涉及三个顺序的步骤：脂质体内吞内化到靶细胞，内体-溶酶体融合，DNA从脂质体中释放并向靶细胞的核移动，以及微管组织显然参与了将导入的外源基因运输到溶酶体内。此外，我们的实验还展示了两种重要的基因转染策略，即使用由生物表面活性物质制成的脂质体和通过激活膜结合的受体分子来利用有效的基因转染。

项目成果

Noguchi, A., Hirashima, N., Nakanishi, M.: "Cationic Choresterol Promotes Gene Transfection Using the Nuclear Localization Signal in Protamine"Pharmaceut. Res.. 19. 933-938 (2002)

Noguchi, A.、Hirashima, N.、Nakanishi, M.：“阳离子胆固醇利用鱼精蛋白中的核定位信号促进基因转染”Pharmaceut。

Dace, A.,Zhao,L.,Park,K.S.,Furuno,T.,Takamura,N.,Nakanishi,M.,West,B., et al.: "Hormone binding induces rapid proteasome-mediated degradation of thyroid hormone receptors."Proc.Natl.Acad.Sci.USA.. 97. 8985-8990 (2000)

Dace, A.、Zhao,L.、Park,K.S.、Furuno,T.、Takamura,N.、Nakanishi,M.、West,B. 等人：“激素结合诱导蛋白酶体介导的甲状腺激素快速降解

Noguchi, S., Hirashima, N., Furuno, T., Nakanishi, M.: "Remarkable induction of apoptsis in cancer cells by a novel cationic liposome complexed with a bcl-2 antisense oligonucleotide"J Control Release. 7. 313-320 (2003)

Noguchi, S.、Hirashima, N.、Furuno, T.、Nakanishi, M.：“与 bcl-2 反义寡核苷酸复合的新型阳离子脂质体显着诱导癌细胞凋亡”J Control Release。

Noguchi, S., Hirashima, N., Furuno, T., Nakanishi, M.: "Remarkable induction of apoptsis in cancer cells by a novel cationic liposome complexed with bcl-2 antisense oligonucleotide"J Control Release. 7. 313-320 (2003)

Noguchi, S.、Hirashima, N.、Furuno, T.、Nakanishi, M.：“与 bcl-2 反义寡核苷酸复合的新型阳离子脂质体显着诱导癌细胞凋亡”J Control Release。

Noguchi,S., Hirashima,N., Furuno,T., Nakanishi,M.: "Remarkable induction of apoptosis in cancer cells by a novel cationic liposome conplexed with a bcl-2 antisense oligonucleotide"J Control Release. 7. 313-320 (2003)

Noguchi,S.、Hirashima,N.、Furuno,T.、Nakanishi,M.：“与 bcl-2 反义寡核苷酸复合的新型阳离子脂质体显着诱导癌细胞凋亡”J Control Release。