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Controlled nonviral gene delivery and expression using stable neural stem cell line transfected with a hypoxia-inducible gene expression system

Authors
 Meng-Lu Liu  ;  Jin Soo Oh  ;  Sung Su An  ;  William A. Pennant  ;  Hyo Jin Kim  ;  So-Jung Gwak  ;  Do Heum Yoon  ;  Keung Nyun Kim  ;  Minhyung Lee  ;  Yoon Ha 
Citation
 JOURNAL OF GENE MEDICINE, Vol.12(12) : 990-1001, 2010 
Journal Title
 JOURNAL OF GENE MEDICINE 
ISSN
 1099-498X 
Issue Date
2010
MeSH
Animals ; Cell Line ; Erythropoietin ; Gene Expression Regulation ; Gene Transfer Techniques ; Genetic Therapy/methods* ; Genetic Vectors ; Hypoxia* ; Mice ; Neural Stem Cells/metabolism ; Neural Stem Cells/transplantation* ; Simian virus 40/genetics ; Transfection/methods*
Abstract
BACKGROUND: Nonviral ex vivo local gene therapy systems consisting of regulated gene expression vectors and cellular delivery platforms represent a novel strategy for tissue repair and regeneration. We introduced a hypoxia-regulated plasmid-based system into mouse neural stem cells (NSCs) as an efficient gene expression and delivery platform for rapid, robust and persistent hypoxic/ischemic-regulated gene expression in the spinal cord. METHODS: A synthetic hypoxia-responsive erythropoietin (Epo) enhancer, the SV40 minimal promoter and the luciferase (Luc) reporter gene were incorporated in a DsRed-expressing double-promoter plasmid for cell lipofection and Zeocin-selection to establish a hypoxia-regulated stable NSC line (NSC-Epo-SV-Luc). A nonhypoxia-regulated stable NSC line (NSC-SV-Luc) was also established as a control. RESULTS: Under the transcriptional regulation of the Epo enhancer, in vitro luciferase expression in NSC-Epo-SV-Luc, but not in NSC-SV-Luc, was sensitively augmented according to the strength and duration of the hypoxic stimulus and was quickly down-regulated to a low basal level after reoxygenation of the hypoxic cells. Furthermore, deoxygenation of the reoxygenated cells clearly enhanced the luciferase activity again. After transplantation into a rat spinal cord injury (SCI) model, only NSC-Epo-SV-Luc showed ischemic injury-specific luciferase expression Notably, the engineered NSC lines kept the neural differentiation potential and retained the hypoxia-regulated luciferase expression after differentiation. CONCLUSIONS: We propose that NSCs engineered with the Epo-SV-therapeutic gene will be valuable for developing a controllable stem cell-mediated nonviral gene therapy for SCI or other central nervous system diseases accompanied with chronic or episodic hypoxic/ischemic stresses
Full Text
http://onlinelibrary.wiley.com/doi/10.1002/jgm.1527/abstract
DOI
10.1002/jgm.1527
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Neurosurgery (신경외과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Medical Research Center (임상의학연구센터) > 1. Journal Papers
1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원) > 1. Journal Papers
Yonsei Authors
Kim, Keung Nyun(김긍년)
Kim, Hyo Jin(김효진)
An, Sung Su(안성수)
Oh, Jin Soo(오진수)
Yoon, Do Heum(윤도흠) ORCID logo https://orcid.org/0000-0003-1452-5724
Ha, Yoon(하윤)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/102595
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