Biosurfactant MEL-A dramatically increases gene transfection via membrane fusion
Biosurfactant MEL-A dramatically increases gene transfection via membrane fusion
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DOI：
10.1016/j.jconrel.2003.10.020
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发表时间：
2004-02-10
影响因子：
10.8
通讯作者：
Nakanishi, M
中科院分区：
文献类型：
--
作者：
Inoh, Y;Kitamoto, D;Nakanishi, M
生物表面活性剂是微生物在非水溶性基质上生长产生的表面活性化合物，具有多种生物活性。本文研究了三种不同的生物表面活性剂，甘露糖脂（MEL）或4-O-[（4'，6'-di-O-acetyl-2'，3'-di-O-alkanoyl）-beta-D-mannopyranosyl] meso-[（MEL-A），4-O-[（6 '-O-乙酰基-2'，3 '-二-O-烷酰基）-β-D-吡喃甘露糖基]内消旋-β-甘露糖基（MEL-B）和4-O-[（4 '-O-乙酰基-2，3'-二-O-烷酰基）-β-D-吡喃甘露糖基]内消旋-β-甘露糖（MEL-C）。MEL-A能提高阳离子脂质体的基因转染效率，而MEL-B和MEL-C则不能。我们还研究了定位的FITC标记的反义DNA（15聚体寡核苷酸;硫代磷酸酯）在靶细胞的激光共聚焦扫描显微镜（CLSM）。FITC标记的反义寡核苷酸暂时位于靶细胞的质膜上，然后将其转移到靶细胞的细胞核中。在MEL-B和MEL-C的情况下，在质膜和细胞核中都没有观察到DNA的这种定位。CLSM图像结果与转染效率一致。这表明MEL-A诱导靶细胞与阳离子脂质体之间的膜融合，显著提高基因转染效率。(C)2003 Elsevier B. V.保留所有权利。
Biosurfactants, which are surface-active compounds produced by microorganisms growing on water-insoluble substrates, have many biological activities. We studied here three different biosurfactants, mannosylerythritol lipid (MEL) or 4-O-[(4',6'-di-O-acetyl-2',3'-di-O-alkanoyl)-beta-D-mannopyranosyl] meso-erythritol (MEL-A), 4-O-[(6'-O-acetyl-2',3'-di-O-alkanoyl)-beta-D-mannopyranosyl] meso-erythritol (MEL-B) and 4-O-[(4'-O-acetyl-2,3'-di-O-alkanoyl)-beta-D-mannopyranosyl] meso-erythritol (MEL-C). MEL-A enhanced the efficiency of gene transfection by cationic liposomes, but MEL-B and MEL-C did not. We also studied the localization of FITC-conjugated antisense DNAs (15-mer oligonucleotides; phosphorothioate) in the target cells by confocal laser scanning microscopy (CLSM). The FITC-conjugated antisense oligonucleotides were temporarily on the plasma membrane of the target cells, thereafter they were transferred into the nucleus of the target cells. In the case of MEL-B and MEL-C, such localization of DNA was not observed both in the plasma membrane and in the nucleus. The results obtained by CLSM images were in good agreement with the transfection efficiency. This suggests that MEL-A induces the membrane fusion between the target cells and the cationic liposomes, accelerating the efficiency of gene transfection dramatically. (C) 2003 Elsevier B.V. All rights reserved.