This CRISPR/Cas9 generated knock-out mutant on the C57BL/6J background of the Chchd10 gene contains a single nucleotide insertion of an A residue within exon 2. These mice may be useful in studies related to mitochondrial bioenergetics and pathogenesis of diseases such as mitochondrial myopathy, motor neuron disease, frontotemporal dementia, Parkinson’s disease, and amyotrophic lateral sclerosis.
Cathleen Lutz, The Jackson Laboratory
The targeted Chchd10 gene encodes a mitochondrial intermembrane protein that is involved with mitochondrial cristae morphology and oxidative phosphorylation. Mutations in the human CHCHD10 gene can cause frontotemporal dementia and amyotrophic lateral sclerosis (ALS).
CRISPR/cas9 endonuclease mediated genome editing of the Chchd10, coiled-coil-helix-coiled-coil-helix domain containing 10, gene was used to introduce a single nucleotide insertion of an A residue within exon 2. This P16fs46ter mutation generates a frameshift that results in termination 46 amino acids after Pro16, creating a knock-out allele. No gene product (protein) is detected by western blot analysis of brain, skeletal muscle and heart tissue from homozygous animals.
Homozygotes are viable and fertile. Skeletal muscle (gastrocnemius) mitochondria from CHCHD10KO mice exhibit decreased ADP-stimulated respiration. Ultrastructural examination of mitochondria reveals electron-dense round structures between sacromeres of the heart right ventricle wall.
Plasmids encoding a signal guide RNA designed to introduce a single nucleotide insertion of an A residue within exon 2 (P16fs46ter) in the Chchd10 gene and the cas9 nuclease were introduced into the cytoplasm C57BL/6J-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to C57BL/6J (Stock No. 000664) to develop the colony. Heterozygotes were intercrossed to generate homozygotes.
|Allele Name||endonuclease-mediated mutation 5, Cathy Lutz|
|Allele Type||Endonuclease-mediated (Null/Knockout)|
|Gene Symbol and Name||Chchd10, coiled-coil-helix-coiled-coil-helix domain containing 10|
|Strain of Origin||C57BL/6J|
|Molecular Note||CRISPR/cas9 endonuclease mediated genome editing was used to introduce a single nucleotide insertion of an A residue within exon 2. This mutation generates a frameshift that results in premature stop codon, creating a knock-out allele. No gene product (protein) is detected by western blot analysis of brain, skeletal muscle and heart tissue from homozygous animals.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the CHCHD10KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #028694 in your Materials and Methods section.