Neural stem cell-specific peroxisome proliferator-activated receptorγknockout mice:breed-ing and genetic identification
10.3969/j.issn.1673-4254.2014.12.12
- VernacularTitle:神经干细胞特异性PPARγ基因敲除小鼠模型的制备与鉴定
- Author:
Qiaoqi WU
1
;
Hongyan ZHANG
;
Zhen WANG
;
Lifang LIN
;
Lu CHEN
;
Xuemin WANG
Author Information
1. 南方医科大学基础医学院神经生物学教研室
- Keywords:
peroxisome proliferator-activated receptorγ;
gene knockout;
Cre-loxp
- From:
Journal of Southern Medical University
2014;(12):1768-1771
- CountryChina
- Language:Chinese
-
Abstract:
Objective To breed neual stem cell-specific peroxisome proliferator-activated receptor γ (PPARγ) knockout mice. Methods Two transgenic mouse models, namely B6.PPARγloxp/loxp and B6.Nestin-Cre were interbred, and the first-generation offsprings were backcrossed with B6.PPARγloxp/loxp to obtain the second-generation mice. Genomic DNA was extracted from the second-generation mice for PCR to amplify the loxp and Cre gene fragments followed by agarose gel electrophoresis to verify their sizes. The mice with the PPARγloxp/loxp.Nestin-Cre (KO) genotype were selected as the neural stem cell-specific knockout PPARγ mice, with B6.PPARγloxp/loxp (loxp) mice as the control. Tissue samples were collected from specific regions of the mouse brain and peripheral tissue for detecting the expression of PPARγ mRNA using RT-PCR and real-time quantitative PCR. Results and Conclusion Genotyping results showed PPARγloxp and Cre bands in the knockout mice, which showed obviously decreased mRNA expression of PPARγ, suggesting successful establishment of neural stem cell-specific PPARγ knockout mice. The two transgenic mice we used were fertile, and their breeding pattern followed the laws of Mendelian inheritance.
