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Comparison of the gene transfer efficiency of mRNA/GL67 and pDNA/GL67 complexes in respiratory cells

Oliwia Andries UGent, Marina De Filette UGent, Joanna Rejman UGent, Stefaan De Smedt UGent, Jo Demeester UGent, Mario Van Poucke UGent, Luc Peelman UGent, Cindy Peleman, Tony Lahoutte and Niek Sanders UGent (2012) MOLECULAR PHARMACEUTICS. 9(8). p.2136-2145
abstract
Complexes between mRNA and GL67:DOPE:DMPE-PEGS000 (GL67) liposomes were formulated and characterized. Subsequently, the in vitro and in vivo expression characteristics of mRNA/GL67 complexes and pDNA/GL67 complexes, each produced at their optimal ratio, were compared in respiratory cells. Transfection of A549 cells with mRNA/GL67 complexes resulted in a much faster expression than after transfection with pDNA/GL67 complexes. The percentage of GFP-positive cells after mRNA and pDNA transfection peaked after 8 and 24 h, respectively. At these time points the percentage of GFP-positive cells was two times higher after mRNA transfection than after pDNA transfection. Furthermore, the efficacy of mRNA/GL67 complexes was independent of the cell cycle. This was in sharp contrast with pDNA/GL67 complexes that caused only a weak expression in nondividing cells. This confirms that the nuclear barrier is a crucial obstacle for pDNA but not for mRNA. Finally, mRNA/GL67 and pDNA/GL67 complexes encoding luciferase were administered intranasally to the lungs of mice. The mRNA/GL67 complexes did not give rise to a measurable luciferase expression in the murine lungs. In contrast, a detectable bioluminescent signal was present in the lungs of mice that received the pDNA/GL67 complexes. We showed that mRNA/GL67 complexes have a lower stability in biological fluids. Consequently, this may be an explanation for their lower performance in vivo.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
gene therapy, mRNA, GL67, lung, nonviral vectors, gene delivery, MESSENGER-RNA, IN-VIVO, MAMMALIAN-CELLS, DELIVERY, DNA, EXPRESSION, STABILITY, RECEPTOR, ACID, LUNG
journal title
MOLECULAR PHARMACEUTICS
Mol. Pharm.
volume
9
issue
8
pages
2136 - 2145
Web of Science type
Article
Web of Science id
000307158000002
JCR category
PHARMACOLOGY & PHARMACY
JCR impact factor
4.57 (2012)
JCR rank
29/259 (2012)
JCR quartile
1 (2012)
ISSN
1543-8384
DOI
10.1021/mp200604h
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2138913
handle
http://hdl.handle.net/1854/LU-2138913
date created
2012-06-11 13:24:45
date last changed
2013-07-08 10:48:49
@article{2138913,
  abstract     = {Complexes between mRNA and GL67:DOPE:DMPE-PEGS000 (GL67) liposomes were formulated and characterized. Subsequently, the in vitro and in vivo expression characteristics of mRNA/GL67 complexes and pDNA/GL67 complexes, each produced at their optimal ratio, were compared in respiratory cells. Transfection of A549 cells with mRNA/GL67 complexes resulted in a much faster expression than after transfection with pDNA/GL67 complexes. The percentage of GFP-positive cells after mRNA and pDNA transfection peaked after 8 and 24 h, respectively. At these time points the percentage of GFP-positive cells was two times higher after mRNA transfection than after pDNA transfection. Furthermore, the efficacy of mRNA/GL67 complexes was independent of the cell cycle. This was in sharp contrast with pDNA/GL67 complexes that caused only a weak expression in nondividing cells. This confirms that the nuclear barrier is a crucial obstacle for pDNA but not for mRNA. Finally, mRNA/GL67 and pDNA/GL67 complexes encoding luciferase were administered intranasally to the lungs of mice. The mRNA/GL67 complexes did not give rise to a measurable luciferase expression in the murine lungs. In contrast, a detectable bioluminescent signal was present in the lungs of mice that received the pDNA/GL67 complexes. We showed that mRNA/GL67 complexes have a lower stability in biological fluids. Consequently, this may be an explanation for their lower performance in vivo.},
  author       = {Andries, Oliwia and De Filette, Marina and Rejman, Joanna and De Smedt, Stefaan and Demeester, Jo and Van Poucke, Mario and Peelman, Luc and Peleman, Cindy and Lahoutte, Tony and Sanders, Niek},
  issn         = {1543-8384},
  journal      = {MOLECULAR PHARMACEUTICS},
  keyword      = {gene therapy,mRNA,GL67,lung,nonviral vectors,gene delivery,MESSENGER-RNA,IN-VIVO,MAMMALIAN-CELLS,DELIVERY,DNA,EXPRESSION,STABILITY,RECEPTOR,ACID,LUNG},
  language     = {eng},
  number       = {8},
  pages        = {2136--2145},
  title        = {Comparison of the gene transfer efficiency of mRNA/GL67 and pDNA/GL67 complexes in respiratory cells},
  url          = {http://dx.doi.org/10.1021/mp200604h},
  volume       = {9},
  year         = {2012},
}

Chicago
Andries, Oliwia, Marina De Filette, Joanna Rejman, Stefaan De Smedt, Jo Demeester, Mario Van Poucke, Luc Peelman, Cindy Peleman, Tony Lahoutte, and Niek Sanders. 2012. “Comparison of the Gene Transfer Efficiency of mRNA/GL67 and pDNA/GL67 Complexes in Respiratory Cells.” Molecular Pharmaceutics 9 (8): 2136–2145.
APA
Andries, O., De Filette, M., Rejman, J., De Smedt, S., Demeester, J., Van Poucke, M., Peelman, L., et al. (2012). Comparison of the gene transfer efficiency of mRNA/GL67 and pDNA/GL67 complexes in respiratory cells. MOLECULAR PHARMACEUTICS, 9(8), 2136–2145.
Vancouver
1.
Andries O, De Filette M, Rejman J, De Smedt S, Demeester J, Van Poucke M, et al. Comparison of the gene transfer efficiency of mRNA/GL67 and pDNA/GL67 complexes in respiratory cells. MOLECULAR PHARMACEUTICS. 2012;9(8):2136–45.
MLA
Andries, Oliwia, Marina De Filette, Joanna Rejman, et al. “Comparison of the Gene Transfer Efficiency of mRNA/GL67 and pDNA/GL67 Complexes in Respiratory Cells.” MOLECULAR PHARMACEUTICS 9.8 (2012): 2136–2145. Print.