Akita Hidetaka, Hama Susumu, Mizuguchi Hiroyuki, Harashima Hideyoshi
Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12 Nishi-6, Kita-ku, Sapporo City 060-0812, Japan.
Yakugaku Zasshi. 2006 Nov;126(11):1047-57. doi: 10.1248/yakushi.126.1047.
For the development of efficient gene vector, intracellular processes such as cellular uptake, endosomal release and nuclear delivery must be overcome. Viruses have also evolved and have developed sophisticated mechanisms for controlling intracellular trafficking for the efficient delivery of their genomes to nuclei in host cells for symbiosis. In the light of these mechanisms, various kinds of artificial devices have been developed to overcome the intracellular barriers. However, in the majority of studies, variation of the transfection activity before and after the modification of devices was evaluated, and intracellular trafficking remained unclear. Therefore, it is understand to recognize which of the intracellular barrier should be intensively improved to enhance the transfection activity. To clarify the rate-limited process in the current non-viral vector, we compared the intracellular trafficking between adenovirus and LipofectAMINE PLUS. As a result, we found that difference of the transfection efficiency between adenovirus and LipofectAMINE PLUS was dominantly derived from the differences on transcription activity. Therefore it is essential to consider the regulation of the intranuclear events to improve the transfection activity of artificial vector.
为了开发高效的基因载体，必须克服诸如细胞摄取、内体释放和核转运等细胞内过程。病毒也在不断进化，并发展出复杂的机制来控制细胞内运输，以便将其基因组高效地递送至宿主细胞的细胞核中实现共生。鉴于这些机制，人们开发了各种人工装置来克服细胞内障碍。然而，在大多数研究中，评估的是装置修饰前后转染活性的变化，而细胞内运输情况仍不清楚。因此，了解应着重改进哪些细胞内障碍以提高转染活性是很有必要的。为了阐明当前非病毒载体中的限速过程，我们比较了腺病毒和LipofectAMINE PLUS之间的细胞内运输情况。结果，我们发现腺病毒和LipofectAMINE PLUS之间转染效率的差异主要源于转录活性的差异。因此，考虑核内事件的调控对于提高人工载体的转染活性至关重要。
