Korean J Bone Metab > Volume 13(2); 2006 > Article
Korean Journal of Bone Metabolism 2006;13(2):119-128.
In vitro and In vivo Transfection Efficiency of Human Osteoprotegerin Gene using Non-Viral Polymer Carriers
폴리머계 유전자 전달체를 이용한 Osteoprotegerin 유전자의 유전자 전달 효율
황창선,강영순,박소영1,임창훈1,김성훈1,윤현구1 홍성란2 한기옥1
성균관의과대학 제일병원 내분비연구실, 내과1, 진단병리과2, 서울대 농생명과학대학3
Abstract
Background
Until now, there are many limitations in the non-viral gene carrier for the clinical applications such as low transfection efficiency and high cytotoxicity. In this study, we examined biodegradable poly (4-vinylimidazole) (P4V) as a non-viral gene carrier and efficacy of in vitro and in vivo transfections. Method: We transfected the HeLa and MC3T3-E1 cell line using P4V, and explored the transfection efficiency and cell cytotoxicity using luciferase and green fluorescent protein expression. PEI (polyethyleneimine) was used as a control gene carrier. For in vivo transfection, 9-week-old Sprague dawley rats were used and divided into control, PEI and P4V groups (n=5). hOPG cDNA encoded pIRES-EYFP construct (200 μg) was mixed with PEI (130 μg) or P4V (230 μg) and DNA/polymer complexes were injected into the tail vein of the rats. After 48h, rats were sacrificed and transfection was observed using confocal microscope and RT-PCR, both at the gene and protein levels in lung and spleen tissue.
Results
The in vitro transfection revealed that P4V had higher transfection efficiency and lower cytotoxicity than PEI. The in vivo experiments confirmed PEI and P4V as non-viral gene carriers. Both PEI and P4V groups showed the expression of the yellow fluorescent protein (YFP), as well as expression of human OPG mRNA in lung and spleen tissue.
Conclusions
P4V was suggested as a more efficient and lower cytotoxic gene carrier than PEI in vitro and in vivo. And further researches are needed to find more effective methods for gene delivery in vivo and to validate the clinical usefulness of the non-viral vector system.
Key Words: Polymer, Transfection, Non-viral gene carrier
TOOLS
METRICS Graph View
  • 825 View
  • 3 Download
Related articles


ABOUT
ARTICLE CATEGORY

Browse all articles >

BROWSE ARTICLES
EDITORIAL POLICY
FOR CONTRIBUTORS
Editorial Office
#1001, Hyundai Kirim Officetel, 42 Seocho-daero 78-gil, Seocho-gu, Seoul 06626, Korea
Tel: +82-2-3473-2231    Fax: +82-70-4156-2230    E-mail: editors.jbm@gmail.com                

Copyright © 2024 by The Korean Society for Bone and Mineral Research.

Developed in M2PI

Close layer
prev next