[Construction of a Mouse Model of CblC Type Methylmalonic Acidemia with W203X Mutation Based on the CRISPR/Cas9 Technology]

Overview

Journal Zhongguo Dang Dai Er Ke Za Zhi

Specialty Pediatrics

Date 2019 Aug 17

PMID 31416510

Citations 1

Authors

Fei Ma

Cong-Cong Shi

Pu-Ping Liang

Si-Tao Li

Xia Gu

Xin Xiao

Hu Hao

Affiliations

Soon will be listed here.

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.

Citing Articles

Versatile enzymology and heterogeneous phenotypes in cobalamin complementation type C disease.

Esser A, Mukherjee S, Derevenkov I, Makarov S, Jacobsen D, Spiekerkoetter U iScience. 2022; 25(9):104981.

PMID: 36105582 PMC: 9464900. DOI: 10.1016/j.isci.2022.104981.

References

Forny P, Schumann A, Mustedanagic M, Mathis D, Wulf M, Nagele N . Novel Mouse Models of Methylmalonic Aciduria Recapitulate Phenotypic Traits with a Genetic Dosage Effect. J Biol Chem. 2016; 291(39):20563-73. PMC: 5034050. DOI: 10.1074/jbc.M116.747717. View

Lerner-Ellis J, Tirone J, Pawelek P, Dore C, Atkinson J, Watkins D . Identification of the gene responsible for methylmalonic aciduria and homocystinuria, cblC type. Nat Genet. 2005; 38(1):93-100. DOI: 10.1038/ng1683. View

Zhou X, Cui Y, Han J . Methylmalonic acidemia: Current status and research priorities. Intractable Rare Dis Res. 2018; 7(2):73-78. PMC: 5982627. DOI: 10.5582/irdr.2018.01026. View

An D, Schneller J, Frassetto A, Liang S, Zhu X, Park J . Systemic Messenger RNA Therapy as a Treatment for Methylmalonic Acidemia. Cell Rep. 2017; 21(12):3548-3558. PMC: 9667413. DOI: 10.1016/j.celrep.2017.11.081. View

Fischer S, Huemer M, Baumgartner M, Deodato F, Ballhausen D, Boneh A . Clinical presentation and outcome in a series of 88 patients with the cblC defect. J Inherit Metab Dis. 2014; 37(5):831-40. DOI: 10.1007/s10545-014-9687-6. View

Peters H, Nefedov M, Sarsero J, Pitt J, Fowler K, Gazeas S . A knock-out mouse model for methylmalonic aciduria resulting in neonatal lethality. J Biol Chem. 2003; 278(52):52909-13. DOI: 10.1074/jbc.M310533200. View

Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna J, Charpentier E . A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012; 337(6096):816-21. PMC: 6286148. DOI: 10.1126/science.1225829. View

Singh K, Evens H, Nair N, Rincon M, Sarcar S, Samara-Kuko E . Efficient In Vivo Liver-Directed Gene Editing Using CRISPR/Cas9. Mol Ther. 2018; 26(5):1241-1254. PMC: 5993986. DOI: 10.1016/j.ymthe.2018.02.023. View

Cong L, Ran F, Cox D, Lin S, Barretto R, Habib N . Multiplex genome engineering using CRISPR/Cas systems. Science. 2013; 339(6121):819-23. PMC: 3795411. DOI: 10.1126/science.1231143. View

10.

Liu M, Yang Y, Chang Y, Chiang S, Lin S, Han L . Mutation spectrum of MMACHC in Chinese patients with combined methylmalonic aciduria and homocystinuria. J Hum Genet. 2010; 55(9):621-6. DOI: 10.1038/jhg.2010.81. View

11.

Nogueira C, Marcao A, Rocha H, Sousa C, Fonseca H, Valongo C . Molecular picture of cobalamin C/D defects before and after newborn screening era. J Med Screen. 2016; 24(1):6-11. DOI: 10.1177/0969141316641149. View

12.

Fowler B, Leonard J, Baumgartner M . Causes of and diagnostic approach to methylmalonic acidurias. J Inherit Metab Dis. 2008; 31(3):350-60. DOI: 10.1007/s10545-008-0839-4. View

13.

Yang Y, Wang L, Bell P, McMenamin D, He Z, White J . A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice. Nat Biotechnol. 2016; 34(3):334-8. PMC: 4786489. DOI: 10.1038/nbt.3469. View

14.

Peters H, Pitt J, Wood L, Hamilton N, Sarsero J, Buck N . Mouse models for methylmalonic aciduria. PLoS One. 2012; 7(7):e40609. PMC: 3392231. DOI: 10.1371/journal.pone.0040609. View