Construction of a mouse model of cblC type methylmalonic acidemia with W203X mutation based on the CRISPR/Cas9 technology.
- Author:
Fei MA
1
;
Cong-Cong SHI
;
Pu-Ping LIANG
;
Si-Tao LI
;
Xia GU
;
Xin XIAO
;
Hu HAO
Author Information
1. Laboratory of Inborn Metabolism, Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou 510655, China. freehaotiger@126.com.
- Publication Type:Journal Article
- MeSH:
Amino Acid Metabolism, Inborn Errors;
Animals;
CRISPR-Cas Systems;
Carrier Proteins;
Heterozygote;
Mice;
Mutation
- From:
Chinese Journal of Contemporary Pediatrics
2019;21(8):824-829
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct a W203X-mutant mouse model of cblC type methylmalonic acidemia based on the CRISPR/Cas9 technology.
METHODS:At first, BLAST was used to compare the conservative nature of the cblC gene and protein sequences in humans and mice, and then, the CRISPR/Cas9 technology was used for microinjection of mouse fertilized eggs to obtain heterozygous F1 mice. Hybridization was performed for these mice to obtain homozygous W203X-mutant mice. The blood level of the metabolite propionyl carnitine (C3) was measured for homozygous mutant mice, heterozygous littermates, and wild-type mice.
RESULTS:The gene and protein sequences of MMACHC, the pathogenic gene for cblC type methylmalonic acidemia, were highly conserved in humans and mice. The homozygous W203X-mutant mice were successfully obtained by the CRISPR/Cas9 technology, and there was a significant increase in C3 in these mice at 24 hours after birth (P<0.001).
CONCLUSIONS:A W203X-mutant mouse model of cblC type methylmalonic acidemia is successfully constructed by the CRISPR/Cas9 technology.
