Metabolic stability of glutaraldehyde cross-linked peptide dna condensates

Abstract

The stability of peptide DNA condensates was examined after introducing glutaraldehyde to cross-link surface amine groups. A 20 amino acid peptide (CWK 18 ) was used to condense DNA into small (70 nm) condensates. The reaction between glutaraldehyde and peptide DNA condensates was indirectly monitored using a fluorescence-based assay to establish reaction completion in 4–5 h when using glutaraldehyde-to-peptide ratios of 1 to 4 mol equiv. Higher levels of glutaraldehyde cross-linking led to significant increases in particle size. The improved stability imparted by glutaraldehyde cross-linking was demonstrated by the increased resistance of DNA condensates to shear stress induced fragmentation. The cross-linked condensates were also significantly more resistant to in vitro metabolism by serum endonucleases. A decrease in the magnitude of transient gene expression was determined for cross-linked DNA condensates which also resulted in a 10-day steady-state expression when cross-linking with 4 mol equiv of glutaraldehyde. The results suggest that cross-linking DNA condensates may provide a means to alter the time course of transient gene expression by inhibiting DNA metabolism.