This CRISPR/Cas9 generated knock-in Tbk1 R228H knock-in mutant of the Tbk1 gene carries a CGG->CAT codon change, resulting in the R228H point mutation. This strain may be useful in studies related to inflammatory response regulation, autophagy, and frontotemporal dementia and/or amyotrophic lateral sclerosis 4.
Cathleen Lutz, The Jackson Laboratory
CRISPR/cas9 endonuclease mediated genome editing of the Tbk1, TANK-binding kinase 1, gene was used to introduce a CGG->CAT codon change (R228H point mutation). The targeted Tbk1 gene encodes a serine/threonine kinase that is involved in inflammatory response regulation and autophagy. Mutations in this gene have been associated with frontotemporal dementia and/or amyotrophic lateral sclerosis 4. Homozygous mice are viable and fertile. As the mice are characterized, we will modify the strain description and add phenotype data.
CRISPR/cas9 endonuclease mediated R228H knock-in point mutation of Tbk1. Plasmids encoding a signal guide RNA designed to introduce the CGG->CAT codon change in the Tbk1 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) for 2 generations to develop the colony.
|Allele Name||endonuclease-mediated 6, Cathy Lutz|
|Allele Type||Endonuclease-mediated (Humanized sequence)|
|Gene Symbol and Name||Tbk1, TANK-binding kinase 1|
|Strain of Origin||C57BL/6J|
|Molecular Note||CRISPR/cas9 genome editing is used to introduce a CGG to CAT codon change resulting in the R228H amino acid substitution. In humans, the R228H substitution is associated with amyotrophic lateral sclerosis.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Tbk1 R228H KI mouse strain in a publication, please cite the originating article(s) and include JAX stock #027983 in your Materials and Methods section.