A method for efficient transduction of miR-483-5p in the kidney of mice.
- Author:
Ying XIA
1
;
Xue-Juan ZHOU
;
Wen-Qing GU
;
Yan-Yan ZHAO
;
Xiao XIAO
;
Xiao-Chun BAI
;
Jun LIU
;
Ming LI
Author Information
- Publication Type:Journal Article
- From: Journal of Southern Medical University 2018;38(2):141-147
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a method for gene delivery in murine renal tissue using lentivirus vector encoding miR-483-5p.
METHODSThirty-five C57BL/6J mice were randomly divided into control group, low-dose treatment group (5 µL each kidney) , and high?dose treatment group (20 µL 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.
RESULTSThe 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).
CONCLUSIONThe 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.