| RRC ID |
42885
|
| Author |
Hama S, Akita H, Iida S, Mizuguchi H, Harashima H.
|
| Title |
Quantitative and mechanism-based investigation of post-nuclear delivery events between adenovirus and lipoplex.
|
| Journal |
Nucleic Acids Res
|
| 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 (E(transcript)) 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 (E(translate)), 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 E(transcript). Moreover, the inferior translation efficiency (E(translate)) 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.
|
| Volume |
35(5)
|
| Pages |
1533-43
|
| Published |
2007-1-1
|
| DOI |
10.1093/nar/gkl1165
|
| PII |
gkl1165
|
| PMID |
17287293
|
| PMC |
PMC1865055
|
| MeSH |
Active Transport, Cell Nucleus
Adenoviridae / genetics*
Cell Line
Cell Nucleus / genetics
Cell Nucleus / metabolism
DNA / analysis
DNA / metabolism
DNA, Viral / analysis
Genetic Vectors
HeLa Cells
Humans
Lipids*
Microscopy, Confocal
Protein Biosynthesis
RNA, Messenger / analysis
RNA, Messenger / metabolism
Transcription, Genetic
Transfection*
Transgenes*
|
| IF |
11.502
|
| Times Cited |
91
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Human and Animal Cells |
HeLa(RCB0007) |
