| The effects of plasmid copy number and sequence context upon transfection efficiency. | |
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MedLine Citation:
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PMID: 14684288 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Wendy E Walker; David J Porteous; A Christopher Boyd |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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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:
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Created Date: 2003-12-19 Completed Date: 2004-09-08 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8607908 Medline TA: J Control Release Country: Netherlands |
Other Details:
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Languages: eng Pagination: 245-52 Citation Subset: IM |
Affiliation:
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Molecular Medicine Centre, Department of Medicine, University of Edinburgh, Western General Hospital, Crewe Road, EH4 2XU, UK. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence COS Cells Cercopithecus aethiops Gene Dosage* Genetic Vectors Plasmids / administration & dosage, genetics, pharmacokinetics* Transfection / methods* |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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