C57BL/6J-Tbk1^em3Lutzy/1J

Stock number: 027080
Product name: Tbk1 G217R KI
Research areas: Immunology, Inflammation and Autoimmunity Research, Neurobiology Research, Research Tools

Description

This CRISPR/Cas9 generated Tbk1 G217R knock-in mutant of the Tbk1 gene carries a G217R knock-in mutation (a GGG->AGG codon change). This strain may be useful in studies related to inflammatory response regulation, autophagy, and frontotemporal dementia and/or amyotrophic lateral sclerosis 4.

Detailed description

Oligo-based CRISPR/cas9 endonuclease-mediated genome editing of the Tbk1 (TANK-binding kinase 1) gene was used to introduce a GGG->AGG codon change (G217R point mutation). The targeted Tbk1 gene encodes a encodes a serine/threonine kinase that is involved in inflammatory response regulation and autophagy. Mutations in this gene are associated with frontotemporal dementia and/or amyotrophic lateral sclerosis 4. Heterozygous mice are viable and fertile, homozygous mice are not viable. As the mice are characterized, we will modify the strain description and add phenotype data.

Development

A mutagenic oligo-based approach was used to generate this strain. Plasmids encoding a signal guide RNA designed to introduce a GGG->AGG codon change (G217R point mutation) in the Tbk1 (TANK-binding kinase 1) 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 DNA sequencing of Tbk1 exon 6 and specific PCR amplified DNA. Mice were further bred to C57BL/6J (Stock# 000664) for 2 generations to develop the colony.

Allele

Symbol: Tbk1^em3Lutzy
Name: endonuclease-mediated mutation 3, Cathy Lutz
MGI accession ID: MGI:6295414
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: Tbk1^G217R
Marker symbol: Tbk1
Marker name: TANK-binding kinase 1
Marker synonyms: 1200008B05Rik; FTDALS4; IIAE8; NAK; T2K
Chromosome: 10
Strain of origin: C57BL/6J
Molecular note: CRISPR/Cas9 genome editing is used to introduce a GGG to AGG codon change resulting in a G217R amino acid substitution. In humans, the G217R substitution is associated with amyotrophic lateral sclerosis.