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Quantitative and mechanism-based investigation of post-nuclear delivery events between adenovirus and lipoplex.

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Title: Quantitative and mechanism-based investigation of post-nuclear delivery events between adenovirus and lipoplex.
Authors: Hama, Susumu Browse this author
Akita, Hidetaka Browse this author
Iida, Shinya Browse this author
Mizuguchi, Hiroyuki Browse this author
Harashima, Hideyoshi Browse this author →KAKEN DB
Issue Date: 2007
Publisher: Oxford University Press
Journal Title: Nucleic Acids Research
Volume: 35
Issue: 5
Start Page: 1533
End Page: 1543
Publisher DOI: 10.1093/nar/gkl1165
PMID: 17287293
Abstract: Quantitative and mechanism-based information on differences in transfection efficiency between viral and non-viral vectors would be highly useful for improving the effectiveness of non-viral vectors. A previous quantitative comparison of intracellular trafficking between adenovirus and LipofectAMINE PLUS (LFN) revealed that the three orders of magnitude lower transfection efficiency of LFN was dominantly rate limited by the post-nuclear delivery process. In the present study, the contribution of transcription and translation processes to the overall differences in the transgene expression efficiency of nucleus-delivered DNA was independently evaluated by quantifying mRNA. As a result, transcription efficiency (Etranscript) of LFN, denoted as transgene expression divided by the amount of nuclear pDNA was about 16 times less than that for adenovirus. Furthermore, translation efficiency (Etranslate), denoted as transfection activity divided by mRNA expression was approximately 460 times less in LFN. Imaging of the decondensed form of DNA by in situ hybridization revealed that poor decondensation efficiency of LFN is involved in the inferior Etranscript. Moreover, the inferior translation efficiency (Etranslate) of LFN was mainly due to electrostatic interactions between LFN and mRNA. Collectively, an improvement in nuclear decondensation and the diminution of the interaction between vector and mRNA is essential for the development of new generations of non-viral vectors.
Type: article
Appears in Collections:薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 原島 秀吉

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