Construction of a testis Elovl4 gene knockout mouse model based on Cre/loxP system.
- Author:
Shisai YANG
1
;
Xuan ZHAO
2
;
Yuhong WANG
2
;
Hongmei ZHENG
2
;
Ting GAN
2
;
Guiming ZHU
2
Author Information
- Publication Type:Journal Article
- Keywords: Cre/loxP; Elovl4; Stra8-Cre; specific knockout; testis
- MeSH: Animals; Disease Models, Animal; Eye Proteins/metabolism*; Female; Gene Knockout Techniques; Integrases; Male; Mammals/metabolism*; Membrane Proteins/metabolism*; Mice; Mice, Knockout; Testis/metabolism*
- From: Chinese Journal of Biotechnology 2022;38(8):2912-2927
- CountryChina
- Language:Chinese
- Abstract: Very long chain polyunsaturated fatty acids (VLC-PUFAs) are unique fatty acids in tissues of mammals such as retina and testis, and the key enzyme of its biosynthesis is very long chain fatty acid elongase 4 (Elovl4). Development of an animal model of tissue-specific knockout of Elovl4 gene is conducive to the in-depth study of the biological function of VLC-PUFAs. Therefore, we constructed Stra8-Cre mice and Elovl4 floxed mice based on Cre/loxP system, and obtained the (Elovl4[flox/+], Stra8-Cre) heterozygous knockout mice by hybridization. Subsequently, female mice were selected to cross with male mice with homozygous Elovl4[flox/flox] to gain homozygous mice (Elovl4[flox/flox], Stra8-Cre) through genotype identification and screening. RT-PCR, qRT-PCR, Western blotting, immunohistochemistry and immunofluorescence techniques were used to detect the knock-out efficiency of Elovl4 in testis. The expression of Elovl4 in testis of both heterozygous and homozygous knockout mice were significantly down-regulated at mRNA and protein levels, but were not affected in other tissues. In summary, we constructed a mouse model with specific knockout of Elovl4 gene in testis, which provides a reliable animal model for studying the effect of VLC-PUFAs on the reproductive function of male mice and the underpinning molecular mechanisms.
