Quantitative and mechanism-based investigation of post-nuclear delivery events between adenovirus and lipoplex.
Quantitative and mechanism-based investigation of post-nuclear delivery events between adenovirus and lipoplex.
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DOI：
10.1093/nar/gkl1165
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发表时间：
2007
影响因子：
14.9
通讯作者：
Harashima H
中科院分区：
文献类型：
--
作者：
Hama S;Akita H;Iida S;Mizuguchi H;Harashima H
关于病毒载体和非病毒载体之间的转染效率差异的定量和基于机制的信息将对提高非病毒载体的有效性非常有用。先前对腺病毒和脂质体(LFN)细胞内转运的定量比较表明，LFN的转染率降低了三个数量级，主要受核后递送过程的限制。在本研究中，转录和翻译过程对核传递DNA转基因表达效率的总体差异的贡献是通过定量mRNA来独立评估的。其结果是，LFN的转录效率(即转基因表达除以核PDNA的量)比腺病毒的转录效率低约16倍。此外，在LFN中，翻译效率(Elate)，表示为转染率除以mRNA表达，大约低460倍。通过原位杂交对去凝集形式的DNA进行成像，发现LFN的去凝集效率较低，这与低转录本有关。此外，LFN的翻译效率较低主要是由于LFN与mRNA之间的静电相互作用。总之，改善核解缩合和减少载体与mRNA之间的相互作用对于开发新一代非病毒载体是至关重要的。
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.
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影响因子：
4.8
作者：
Martins, RP;Ostermeier, GC;Krawetz, SA
通讯作者：
Krawetz, SA
影响因子：
10.8
作者：
Moriguchi, R;Kogure, K;Harashima, H
通讯作者：
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影响因子：
3.7
作者：
HATFIELD, L;HEARING, P
通讯作者：
HEARING, P
影响因子：
3.2
作者：
Keller, M;Harbottle, RP;Miller, AD
通讯作者：
Miller, AD
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2.7
作者：
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通讯作者：
Wadell, G