PMID- 19692581
OWN - NLM
STAT- MEDLINE
DCOM- 20091214
LR  - 20211020
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 37
IP  - 18
DP  - 2009 Oct
TI  - Oligopeptide-mediated gene transfer into mouse corneal endothelial cells: 
      expression, design optimization, uptake mechanism and nuclear localization.
PG  - 6276-89
LID - 10.1093/nar/gkp651 [doi]
AB  - 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.
FAU - Seow, Wei Yang
AU  - Seow WY
AD  - Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, Singapore 
      138669, Singapore.
FAU - Yang, Yi-Yan
AU  - Yang YY
FAU - George, Andrew J T
AU  - George AJ
LA  - eng
GR  - Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090819
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Indoleamine-Pyrrole 2,3,-Dioxygenase)
RN  - 0 (Oligopeptides)
RN  - 9007-49-2 (DNA)
RN  - 94ZLA3W45F (Arginine)
SB  - IM
MH  - Animals
MH  - Arginine/chemistry
MH  - Cell Nucleus/chemistry
MH  - Cells, Cultured
MH  - DNA/analysis
MH  - Endothelium, Corneal/*metabolism
MH  - Gene Expression
MH  - Genes, Reporter
MH  - Indoleamine-Pyrrole 2,3,-Dioxygenase/metabolism
MH  - Mice
MH  - Oligopeptides/*chemistry
MH  - *Transfection
PMC - PMC2764440
EDAT- 2009/08/21 09:00
MHDA- 2009/12/16 06:00
PMCR- 2009/08/19
CRDT- 2009/08/21 09:00
PHST- 2009/08/21 09:00 [entrez]
PHST- 2009/08/21 09:00 [pubmed]
PHST- 2009/12/16 06:00 [medline]
PHST- 2009/08/19 00:00 [pmc-release]
AID - gkp651 [pii]
AID - 10.1093/nar/gkp651 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2009 Oct;37(18):6276-89. doi: 10.1093/nar/gkp651. Epub 2009 
      Aug 19.