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The Role of a Microscopic Colloidally Stabilized Phase in Solubilizing Oligoamine-Condensed DNA Complexes

Vladimir S. Trubetskoy^*, Jon A. Wolff and Vladimir G. Budker

^* Mirus Corporation, Madison, Wisconsin 53719; and Departments of Pediatrics and Medical Genetics, Waisman Center, University of Wisconsin, Madison, Wisconsin 53705

Correspondence: Address reprint requests to Dr. Vladimir S. Trubetskoy, Mirus Corp., 505 S. Rosa Rd., Madison, WI 53719. Tel.: 608-441-2832; Fax: 608-441-2849; E-mail: vladimirt{at}genetransfer.com.

DNA complexes of spermine and spermidine become resolubilized at very high concentrations of the oligoamine. It has been postulated that high oligoamine concentrations shift the DNA from the globule back to the coil phase. The present study indicates that DNA resolubilization at high concentrations of spermine and spermidine is explained by formation of small particles of condensed DNA that cannot be precipitated by centrifugation. The fact that DNA stays condensed during resolubilization was confirmed using a relatively new condensation assay and three independent microscopic techniques. A considerable portion of DNA was found to be in particles with diameter <100 nm. Formation of such small particles is likely to be caused by colloidal forces. The ability to form small, condensed DNA particles in solutions that contain high concentrations of oligocation should aid in the design of synthetic DNA vectors for gene transfer and gene therapy and in the handling of DNA for diagnostic studies.