| Optimization of cell surface binding enhances efficiency and specificity of molecular conjugate gene delivery. | |
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MedLine Citation:
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PMID: 9774415 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Molecular conjugates, or polyplexes, are promising synthetic vectors for targeted, in vivo gene delivery, if their efficiency can be improved. Gaining mechanistic information on conjugate gene delivery can potentially yield significant improvements in transfer efficiency by revealing barriers to conjugate transfer from the cell surface to the nucleus. We have developed an experimental system that employs epidermal growth factor as the ligand to direct delivery of DNA encoding the green fluorescent protein to mouse fibroblasts. We report here that the initial step of delivery, binding of the conjugate to the cell surface, is a barrier to gene transfer. We examined the effects of conjugate charge, ligand cross-linker spacer length, and ligand valency on polyplex cell surface binding, internalization, and gene transfer. We find that delivery is both efficient and specific only within a relatively narrow window of conjugate charge, results that correlate with binding and internalization of radiolabeled conjugate. In addition, increasing the cross-linker length can improve binding affinity and delivery. Finally, there is a significant optimum in gene delivery as a function of ligand valency, due to saturation of receptor binding and internalization. Optimizing parameters that affect surface binding therefore improves the efficiency and specificity of molecular conjugate gene delivery. |
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Authors:
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D V Schaffer; D A Lauffenburger |
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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: The Journal of biological chemistry Volume: 273 ISSN: 0021-9258 ISO Abbreviation: J. Biol. Chem. Publication Date: 1998 Oct |
Date Detail:
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Created Date: 1998-11-12 Completed Date: 1998-11-12 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 28004-9 Citation Subset: IM |
Affiliation:
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Department of Chemical Engineering and Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. dschaffer@ems.salk.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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3T3 Cells Animals Biotinylation Cell Membrane / metabolism Down-Regulation Epidermal Growth Factor / metabolism* Gene Targeting / methods* Green Fluorescent Proteins Ligands Luminescent Proteins / genetics* Mice Protein Binding Receptor, Epidermal Growth Factor / metabolism |
| Chemical | |
Reg. No./Substance:
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0/Ligands; 0/Luminescent Proteins; 147336-22-9/Green Fluorescent Proteins; 62229-50-9/Epidermal Growth Factor; EC 2.7.10.1/Receptor, Epidermal Growth Factor |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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