Characterizing DNA condensation and conformational changes in organic solvents
| Autor: | Yen K. Luu, Dehai Liang, Fuyou Ke, Michael Hadjiargyrou |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Chemistry/Macromolecular Chemistry
Gel electrophoresis of nucleic acids lcsh:Medicine Gene delivery DNA condensation chemistry.chemical_compound Plasmid Microscopy Electron Transmission Calcium chloride transformation Animals Denaturation (biochemistry) Biochemistry/Macromolecular Chemistry lcsh:Science Electrophoresis Agar Gel Multidisciplinary Oligonucleotide Chemical Biology/Macromolecular Chemistry Circular Dichroism lcsh:R DNA Biochemistry chemistry Biophysics Solvents Nucleic Acid Conformation lcsh:Q Cattle Spectrophotometry Ultraviolet Plasmids Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 5, Iss 10, p e13308 (2010) |
| ISSN: | 1932-6203 |
| Popis: | Organic solvents offer a new approach to formulate DNA into novel structures suitable for gene delivery. In this study, we examined the in situ behavior of DNA in N, N-dimethylformamide (DMF) at low concentration via laser light scattering (LLS), TEM, UV absorbance and Zeta potential analysis. Results revealed that, in DMF, a 21bp oligonucleotide remained intact, while calf thymus DNA and supercoiled plasmid DNA were condensed and denatured. During condensation and denaturation, the size was decreased by a factor of 8-10, with calf thymus DNA forming spherical globules while plasmid DNA exhibited a toroid-like conformation. In the condensed state, DNA molecules were still able to release the counterions to be negatively charged, indicating that the condensation was mainly driven by the excluded volume interactions. The condensation induced by DMF was reversible for plasmid DNA but not for calf thymus DNA. When plasmid DNA was removed from DMF and resuspended in an aqueous solution, the DNA was quickly regained a double stranded configuration. These findings provide further insight into the behavior and condensation mechanism of DNA in an organic solvent and may aid in developing more efficient non-viral gene delivery systems. |
| Databáze: | OpenAIRE |
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