A method for efficient transduction of miR-483-5p in the kidney of mice

XIA Ying; ZHOU Xuejuan; GU Wenqing; ZHAO Yanyan; XIAO Xiao; BAI Xiaochun; LIU Jun; LI Ming

doi:10.3969/j.issn.1673-4254.2018.02.04

Abstract：Objective To establish a method for gene delivery in murine renal tissue using lentivirus vector encoding miR-483-5p. Methods Thirty-five C57BL/6J mice were randomly divided into control group, low-dose treatment group (5μL each kidney) , and high-dose treatment group (20μL each kidney), and in the latter two groups, the lentivirus vector encoding miR-483-5p were injected in the renal cortex. The tissue samples were collected at 7 and 21 days after the injection. A transgenic mouse model with inducible systemic overexpression of miR-483-5p was established in TG483 mice. The Cre-loxp system was used to create a mouse model with renal tubule-specific expression of miR-483-5p. The levels of BUN in the mice were detected and HE staining and fluorometric TUNEL assay were used to observe the morphological changes of the kidneys;real-time qPCR was used to detect miR-483-5p expression in the renal cortex. Results The mice with overexpression of miR-483-5p had normal renal function without obvious pathological changes or apoptosis in the renal tissue. Renal cortex injection of 20 μL lentivirus resulted in obviously increased level of miR-483-5p at 21 days (1.2 ± 0.43 vs 8.6 ± 1.09, P<0.001). miR-483-5p showed a low expression (0.9 ± 0.09 vs 1.7 ± 0.19, P<0.05) in TG483 mice and a high expression in the kidney of the transgenic mice established using the Cre-loxp system (1.6 ± 1.13 vs 12.36 ± 3.89, P<0.05). Conclusion The transgenic mice with renal tubule-specific expression of miR-483-5p show normal renal function, and this model facilitates further study of the role of miR-483-5p in the kidney.