Various properties of polymeric carriers improved the transfection efficiency.
Various properties of polymeric carriers improved the transfection efficiency.
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DOI：
10.1093/nass/nrl166
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发表时间：
2006-11
期刊：
影响因子：
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通讯作者：
T. Hashimoto;A. Mahara;A. Kobori;A. Murakami;T. Yamaoka
中科院分区：
文献类型：
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作者：
T. Hashimoto;A. Mahara;A. Kobori;A. Murakami;T. Yamaoka
为了开发新型高效基因载体，我们一直关注基因转移系统各个步骤中转基因在细胞核中的转录。迄今为止，改善转录因子转基因识别的最佳载体特性尚未阐明。在本研究中，我们利用显微注射技术建立了一种新的转基因核内转录效率评价体系。将由不同分子量的多肽组成的聚合复合物显微注射到COS-1细胞的细胞质或细胞核中，并评估载体特性（例如分子量）与核内转录效率之间的关系。活细胞中转基因转录的分子量（Mw）依赖性已成功定量，发现低Mw聚合物不会抑制转录，但高Mw聚合物几乎不允许转录。有趣的是，聚精氨酸 (PR) 和聚赖氨酸 (PK) 的转录效率几乎相同，尽管 PR 被广泛报道为比 PK 更有效的基因载体。我们将讨论 PR/DNA 和 PK/DNA 复合物的各种特性和细胞内运输的差异。
In order to develop novel efficient gene carriers, we have been focusing on the transcription of transgene in the nucleus among various steps in the gene transfer system. Optimal carrier properties for improving the transgene recognition by transcription factors have not been clarified so far. In the present study, we established a novel evaluation system for the intranuclear transcription efficiency of the transgene using microinjection technique. Polyplexes composed of polypeptides with different molecular weights were microinjected into the cytoplasm or nucleus of COS-1 cells, and the relationship between the carrier properties, such as molecular weight, and the intranuclear transcription efficiency was evaluated. The molecular weight (Mw) dependency of the transgene transcription in the living cells was successfully quantified, and the low Mw polymers were found not to suppress the transcription but high Mw polymers allowed almost no transcription. Interestingly, the transcription efficiencies of poly(arginine) (PR) and poly(lysine) (PK) were almost same although the PR is widely reported to be more efficient gene carrier than PK. The difference in the various properties and intracellular trafficking of PR/DNA and PK/DNA polyplexes will be discussed.