Skip Navigation


Nucleic Acids Research Advance Access originally published online on August 19, 2009
Nucleic Acids Research 2009 37(18):6276-6289; doi:10.1093/nar/gkp651
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (2356K) Freely available
Right arrow Screen PDF (1314K) Freely available
Right arrow Supplementary Data
Right arrowOA All Versions of this Article:
37/18/6276    most recent
gkp651v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Seow, W. Y.
Right arrow Articles by George, A. J. T.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Seow, W. Y.
Right arrow Articles by George, A. J. T.
Related Collections
Right arrow DNA transfer
Right arrow Miscellaneous/other
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2009, Vol. 37, No. 18 6276-6289
© The Author 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses?by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Synthetic Biology and Chemistry

Oligopeptide-mediated gene transfer into mouse corneal endothelial cells: expression, design optimization, uptake mechanism and nuclear localization

Wei Yang Seow1,2, Yi-Yan Yang1,* and Andrew J. T. George2,*

1Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, Singapore 138669, Singapore and 2Department of Immunology, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK

*To whom correspondence should be addressed. Tel: +44 20 8383 1475; Fax: +44 20 8383 2788; Email: a.george{at}imperial.ac.uk

Correspondence may also be addressed to Yi-Yan Yang. Tel: +65 6824 7106; Fax: +65 6478 9084; Email: yyyang{at}ibn.a-star.edu.sg

Received June 21, 2009. Revised July 21, 2009. Accepted July 21, 2009.

Gene transfer to the corneal endothelium has potential in preventing corneal transplant rejection. In this study, we transfected mouse corneal endothelial cells (MCEC) with a class of novel arginine-rich oligopeptides. The peptides featured a tri-block design and mediated reporter gene expression in MCEC more efficiently than the commercial polyethylenimine standard. The functionality of each block was demonstrated to critically influence the performance of the peptide. Results from confocal imaging and flow cytometry then showed that energy-dependent endocytosis was the dominant form of uptake and multiple pathways were involved. Additionally, uptake was strongly dependent on interactions with cell-surface heparan sulphate. Fluorescence resonance energy transfer studies revealed that the peptide/DNA entered cells as an associated complex and some will have dissociated by 8.5 h. Large-scale accumulation of uncondensed DNA within the nucleus can also be observed by 26 h. Finally, as a proof of biological relevance, we transfected MCEC with plasmids encoding for the functional indoleamine 2,3-dioxygenase (IDO) enzyme. We then demonstrated that the expressed IDO could catalyse the degradation of L-tryptophan, which in turn suppressed the growth of CD4+ T-cells in a proliferation assay.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.