Vol.6 No.3 (September 2016)
Hydrodynamic Transfection of Double Reporter Genes into Mouse Liver
陈 丽 ：广州医科大学附属第二医院，广东 广州
肖 东 ：南方医科大学肿瘤研究所，广东 广州；南方医科大学实验动物中心暨比较医学研究所，广东 广州
Objective: Set up the system for hydrodynamic transfection of double reporter genes [green fluo-rescent protein (GFP) and firefly luciferase (Luc)] into mouse liver. Methods: To in vitro verify the functionality of pcEF.luc-IRES-GFP, human hepatocellular carcinoma 7721 cells were transiently transfected with pcEF.luc-IRES-GFP, followed by GFP and Luc assay. Hydrodynamic transfection was employed to deliver pcEF.luc-IRES-GFP into mice via tail-vein injection, followed by in vivo and ex vivo imaging to detect Luc and GFP. Moreover, immunohistochemistry (IHC) were performed to detect GFP expression in mouse liver cells. Results: The expression of GFP and Luc was detected in 7721 cell transiently transfected by pcEF.luc-IRES-GFP. Luc signals were detected in liver by in vivo bioluminescence imaging, and both Luc and GFP were detected in isolated liver by in vivo bioluminescence and fluorescence imaging. IHC revealed the GFP-positive liver cells of mice hydrodynamically transfected via tail-vein injection of pcEF.luc-IRES-GFP. Conclusions: We successfully deliver double reporter genes (GFP and luciferase) into mouse liver by hydrodynamic transfection, which lays a solid foundation for further research.
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