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Nucleic Acids Research, 2002, Vol. 30, No. 6 1338-1345
© 2002 Oxford University Press

Optimal transfection with the HK polymer depends on its degree of branching and the pH of endocytic vesicles

Qing-Rong Chen, Lei Zhang, Paul W. Luther1 and A. James Mixson*

Department of Pathology, University of Maryland Baltimore, MSTF Building, 10 South Pine Street, Baltimore, MD 21201, USA and 1Department of Physiology, University of Maryland Baltimore, Howard Hall, 660 West Redwood Street, Baltimore, MD 21201, USA

We have recently reported that liposomes in combination with histidine (HK)-containing polymers enhanced the expression of luciferase in transfected cells. In transformed or malignant cell lines, branched HK polymers (combined with liposome carriers) were significantly more effective than the linear HK polymer in stimulating gene expression. In the current study, we found that the linear HK polymer enhanced gene expression in primary cell lines more effectively than the branched polymers. The differences in the optimal carrier (linear versus branched) were not due to initial cellular uptake, size of the complexes or level of gene expression. There was, however, a strong association between the optimal type of HK polymer and the pH of endocytic vesicles (P = 0.0058). By altering the percentage of histidines carrying a positive charge, the endosomal pH of a cell may determine the amount of DNA released from the linear or branched HK polymer. In the two cell lines in which the linear HK was the optimal polymer, the endocytic vesicles were strongly acidic with a pH of <5.0. Conversely, in the four cell lines in which the branched polymers were optimal transfection agents, the pH of endocytic vesicles was >6.0. Furthermore, binding data support the relationship between DNA release from the optimal HK polymer and endosomal pH. The interplay between optimal HK polymers and the endosomal pH may lead to improved gene-delivery polymers tailored to a particular cell.

* To whom correspondence should be addressed. Tel: +1 410 706 3223; Fax: +1 410 706 8414; Email: amixson{at}umaryland.edu


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