Efficient gene delivery using reconstituted chromatin enhanced for nuclear targeting Kylie M. Wagstaff * , Jun Y. Fan ‡ , Michelle A. De Jesus * , David J. Tremethick ‡ and David A. Jans * ,† ,1 * Nuclear Signalling Laboratory, Department of Biochemistry and Molecular Biology, and † Australian Research Council Centre of Excellence for Biotechnology and Development, Monash University, Clayton, Australia; and ‡ Chromatin Transcription and Regulation Laboratory, John Curtin School of Medical Research, Australian National University, Canberra, Australia 1 Correspondence: Nuclear Signalling Laboratory, Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800 Australia. E-mail: david.jans@med.monash.edu.au Nonviral gene delivery is hampered by difficulties associated with transporting negatively charged DNA through the cell membrane and, more importantly, the nuclear envelope of target cells. Here we show for the first time that chromatin reconstituted with histone H2B proteins optimized for nuclear targeting can be used as an efficient means to deliver DNA to the nucleus of intact living mammalian cells, resulting in high levels of transgene expression that were ∼6-fold more than those achieved by commercial liposomal preparations. The high efficiency is due in part to DNA condensation and protection against degradation in the reconstituted chromatin, as well as its ability to interact with high affinity with the importin proteins of the cellular nuclear import machinery. "Chromofection," gene delivery by protein transduction using chromatin enhanced for nuclear targeting represents an efficient means to deliver DNA to a wide variety of cell types, with the potential to treat complex genetic disorders.—Wagstaff, K. M., Fan, J. Y., De Jesus, M. A., Tremethick, D. J., Jans, D. A. Efficient gene delivery using reconstituted chromatin enhanced for nuclear targeting. Key Words: gene therapy • histone • protein transduction • NLS
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Efficient gene delivery using reconstituted chromatin enhanced for nuclear targeting
Wagstaff, Kylie M.; Fan, Jun Y.; De Jesus, Michelle A.; Tremethick, David J.; Jans, David A.
Follow The FASEB Journal
, Volume 22 (7): 2232
Fed of American Socs for Experimental Biology – Jul 1, 2008
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