Loosening of DNA/polycation complexes by synthetic polyampholyte to improve the transcription efficiency: effect of charge balance in the polyampholyte.
High mobililty group proteins are amphoteric nuclear proteins that are known to unfold chromatin to stimulate transcription. To mimic their structures, we synthesized the novel polyethylene glycol (PEG) derivatives, PEG-ACs, consisting of both amino- and carboxyl-pendants in various ratios, and their loosening and transcription-improving activity on the DNA complex was examined. Fluorescence anisotropy measurement revealed that anionic PEG-ACs with more carboxyls than amines could efficiently loosen the DNA/polyethyleneimine complex. Those anionic PEG-ACs showing a loosening effect on the DNA complex evidently increased the transcription rate to >20 times higher than that of the original complex, probably owing to the facilitated approach of transcriptional factors to the DNA segments in the loosened complexes. The complexes with anionic PEG-ACs also showed improved transgene expression level on the cultured cells, indicating the effectiveness of improving transcriptional activity to attain a high extragene expression by the plasmid complex. The loosening mechanism of DNA/polycation complexes was investigated with a simplified model via Monte Carlo simulation to discern the difference in the presence of cationic polyampholytes, anionic polyampholytes, and polyanions.
Yoshihara
Chieko
C
Otsuma Women's University Graduate School, Sanbancho, Chiyoda-ku, Tokyo, Japan.
Shew
Chwen-Yang
CY
Ito
Tomoko
T
Koyama
Yoshiyuki
Y
D000838:Anions; D016880:Anisotropy; D001703:Biophysics / Q000379:methods*; D002412:Cations; D004247:DNA / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D008957:Models, Genetic; D009010:Monte Carlo Method; D010957:Plasmids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D013050:Spectrometry, Fluorescence / Q000379:methods; D055672:Static Electricity; D014158:Transcription, Genetic; D019076:Transgenes
D000838:Anions; D002412:Cations; D011092:Polyethylene Glycols; D004247:DNA