Strength of conjugate binding to plasmid DNA affects degradation rate and expression level in vivo

Autor: Arjang Amini, Joanne M Fabrycki, Quynh-Chi Phan, Dennis J. Carlo, Patrick Pezzoli, Christopher C. Coffin, Charles P. Lollo, Mariusz G. Banaszczyk, Patricia M. Mullen, Alison T. Carlo, Dongpei Wu
Rok vydání: 2000
Předmět:
Zdroj: Biochimica et Biophysica Acta (BBA) - General Subjects. 1523:103-110
ISSN: 0304-4165
DOI: 10.1016/s0304-4165(00)00104-5
Popis: In vitro assays have demonstrated the capability of poly- L -lysine to protect plasmid DNA from serum nucleases and cellular lysates. Our purpose was to evaluate the stability and potency of poly- L -lysine–DNA polyplexes after intravenous injection into mice. Polyplexes consisted of 32 P-radiolabeled plasmid DNA complexed with poly- L -lysine at specified charge ratios. Variations in conjugate hydrophobicity and levels of modification with polyethylene glycol were investigated. Our results show that, in contrast to in vitro studies, the systemically administered polyplexes exhibited marked DNA degradation in the vascular compartment within 5 min. Substitution of poly- L -lysine ϵ-amino sites with polyethylene glycol or hydrocarbon chains resulted in faster degradation even when complexed at higher charge (±) ratios. Use of excess cationic charge in the polyplexes (±2.5) diminished degradation rates only slightly. An analysis was made of the strength of the poly- L -lysine:DNA interaction by competition with poly-aspartic acid. Polyplexes with the strongest binding between conjugate and DNA in the competition assay were also the most stable in blood. However, tighter binding was not enough to fully protect the polyplex in vivo and polyplex DNA was substantially degraded within 10 min. Increased polyplex stability did not correlate with improved in vivo transfection efficiency.
Databáze: OpenAIRE
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