Document Detail

The effects of plasmid copy number and sequence context upon transfection efficiency.
MedLine Citation:
PMID:  14684288     Owner:  NLM     Status:  MEDLINE    
It is known that large P1 artificial chromosome (PAC) vectors exhibit reduced transfection efficiency in comparison to small plasmid vectors. We investigated the dynamics of this effect, by comparing expression from a small plasmid (4.7 kb) and a PAC vector (111 kb) containing the Enhanced Green Fluorescent Protein (EGFP) reporter gene under the control of a P(CMV) promoter. EGFP expression was detected by fluorescence activated cell sorting (FACS). We found that the lower transfection efficiency of PAC vectors represents both a smaller percentage of cells expressing the transgene, and a lower level of expression per cell. We have shown that the lower number of plasmid molecules administered per cell in a PAC transfection does not explain this effect, and that this effect does not act in trans. Surprisingly, dilution of a reporter construct with an irrelevant plasmid did not appear to compromise transfection efficiency; in fact, a dilution of 1/10 slightly enhanced transfection. Therefore, it seems that the plasmid content of a liposome-DNA complex need not be 100% reporter construct for optimum transfection efficiency. This discovery has potential practical utility in a number of applications.
Wendy E Walker; David J Porteous; A Christopher Boyd
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of controlled release : official journal of the Controlled Release Society     Volume:  94     ISSN:  0168-3659     ISO Abbreviation:  J Control Release     Publication Date:  2004 Jan 
Date Detail:
Created Date:  2003-12-19     Completed Date:  2004-09-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8607908     Medline TA:  J Control Release     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  245-52     Citation Subset:  IM    
Molecular Medicine Centre, Department of Medicine, University of Edinburgh, Western General Hospital, Crewe Road, EH4 2XU, UK.
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MeSH Terms
Base Sequence
COS Cells
Cercopithecus aethiops
Gene Dosage*
Genetic Vectors
Plasmids / administration & dosage,  genetics,  pharmacokinetics*
Transfection / methods*

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