Incorporation of Viral Glycoprotein VSV-G Improves the Delivery of DNA by Erythrocyte Ghost into Cells Refractory to Conventional Transfection.
The objective of this study was to formulate a novel gene delivery system based on the erythrocyte ghost (EG) integrated with fusogenic viral glycoprotein vesicular stomatitis virus glycoprotein G (VSV-G). VSV-G proteins were harvested as condition medium of Ad293 cells carrying a VSV-G transgene and then incorporated into EG. Plasmid DNA was condensed by various transfection reagents. A luciferase expression construct (pGL3-control) and a DsRed expression cassette (pCMV-DsRed) were used to evaluate the delivery efficiency of DNA/EG/VSV-G complexes. VSV-G proteins could be incorporated into EG in static incubation under acidic conditions as evidenced by the Western blot analysis. Condensed plasmid DNA was bound mostly to the outer surface of EG, which could be detected by electromicroscopy and measured by electrophoresis. EG/VSV-G complexes stimulated the delivery of pGL3-control into Ad293 cells significantly with the luciferase activity increased about 4-fold as compared to that of the control. The delivery of pCMV-DsRed was also enhanced with the percentage of DsRed-positive Ad293 cells increased from 55 % to about 80 %. Moreover, the transfection efficiency in 3T3, HeLa, INS-1, and bone marrow stem cell (BMSC) cells increased about 2-3-fold. Finally, confocal microscopy analysis showed that incorporation of VSV-G significantly enhanced the endocytosis of EG into target cells. In the present study, a novel type of non-viral DNA delivery vehicle consisting of EG and fusogenic VSV-G proteins was formulated, which showed superior transfection efficiency even in cells resistant to classical transfection.
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China.
Xin
X
Liu
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China.
Yun-Pan
YP
Li
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China.
Zhen-Min
ZM
Zhong
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China.
Hui-Qi
HQ
Tan
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China.
Hao-Peng
HP
Lin
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China.
Shao-Jun
SJ
Chen
Laboratory of Cell Senescence, Shantou University Medical College, Shantou, Guangdong, 515041, China.
Yu-Cai
YC
Fu
Department of Endocrinology, the First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, 515041, China.
Wen-Can
WC
Xu
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, 515063, China. chijuwei@stu.edu.cn.
Chi-Ju
CJ
Wei
D016475:3T3 Cells; D000818:Animals; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D004910:Erythrocyte Membrane / Q000235:genetics*; D024861:Genetic Enhancement / Q000379:methods*; D006023:Glycoproteins / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D014162:Transfection / Q000379:methods*
D006023:Glycoproteins; D004247:DNA
DNA delivery; Erythrocyte ghost; Luciferase activity; VSV-G