Hokkaido University Collection of Scholarly and Academic Papers >
Faculty of Pharmaceutical Sciences >
Peer-reviewed Journal Articles, etc >
Quantitative analysis of condensation/decondensation status of pDNA in the nuclear sub-domains by QD-FRET
Title: | Quantitative analysis of condensation/decondensation status of pDNA in the nuclear sub-domains by QD-FRET |
Authors: | Shaheen, Sharif M. Browse this author | Akita, Hidetaka Browse this author | Yamashita, Atsushi Browse this author | Katoono, Ryo Browse this author | Yui, Nobuhiko Browse this author | Biju, Vasudevanpillai Browse this author | Ishikawa, Mitsuru Browse this author | Harashima, Hideyoshi Browse this author |
Issue Date: | Apr-2011 |
Publisher: | Oxford University Press |
Journal Title: | Nucleic Acids Research |
Volume: | 39 |
Issue: | 7 |
Start Page: | e48 |
Publisher DOI: | 10.1093/nar/gkq1327 |
Abstract: | Recent studies indicate that controlling the nuclear decondensation and intra-nuclear localization of plasmid DNA (pDNA) would result in an increased transfection efficiency. In the present study, we established a technology for imaging the nuclear condensation/decondensation status of pDNA in nuclear subdomains using fluorescence resonance energy transfer (FRET) between quantum dot (QD)-labeled pDNA as donor, and rhodamine-labeled polycations as acceptor. The FRET-occurring pDNA/polycation particle was encapsulated in a nuclear delivery system; a tetra-lamellar multifunctional envelope-type nano device (T-MEND), designed to overcome the endosomal membrane and nuclear membrane via step-wise fusion. Nuclear subdomains (i.e. heterochromatin and euchromatin) were distinguished by Hoechst33342 staining. Thereafter, Z-series of confocal images were captured by confocal laser scanning microscopy. pDNA in condensation/decondensation status in heterochromatin or euchromatin were quantified based on the pixel area of the signals derived from the QD and rhodamine. The results obtained indicate that modulation of the supra-molecular structure of polyrotaxane (DMAE-ss-PRX), a condenser that is cleaved in a reductive environment, conferred euchromatin-preferred decondensation. This represents the first demonstration of the successful control of condensation/decondensation in specific nuclear sub-domain via the use of an artificial DNA condenser. |
Type: | article |
URI: | http://hdl.handle.net/2115/45389 |
Appears in Collections: | 薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
|
Submitter: 原島 秀吉
|