C57BL/6J-Tardbp^em9Lutzy Snca^em2Lutzy Mapt^em2Lutzy/KmsoJ

Stock number: 034028
Product name: Tardbp^G348C ; Snca^A30P ; Mapt^P290L
Strain synonyms: Tardbp G348C KI ; Snca A30P ; Mapt V289V P290L
Research areas: Developmental Biology Research, Mouse/Human Gene Homologs, Neurobiology Research

Description

The Tardbp^G348C ; Snca^A30P ; Mapt^P290L triple mutant strain harbors several mutations associated with human neurodegenerative disease.
Tardbp^G348C encodes a mutation found in Lou Gehrig's disease (Amyotrophic lateral sclerosis; ALS) patients.
Snca^A30P mimics a human mutation associated with synucleinopathies such as Parkinson's disease and multiple system atrophy, as well as Lewy body dementia.
Mapt^P290L is equivalent to the human MAPT P301L mutation identified in frontotemporal dementia with parkinsonism.

Detailed description

Stock No. 034028 Tardbp^G348C ; Snca^A30P ; Mapt^P290L (aka, Tardbp G348C KI ; Snca A30P ; Mapt V289V P290L) is a triple mutant strain harboring several mutations associated with human neurodegenerative disease. Mice homozygous for all three mutant alleles are viable and fertile [January 2025]. If additional characterization of these mice is performed, we may modify the strain description accordingly. Each allele is described below.

The Tardbp^G348C allele encodes a mutation found in Lou Gehrig's disease (Amyotrophic lateral sclerosis; ALS) patients. The endogenous Tardbp gene encodes the DNA and RNA-binding protein TDP-43. Ubiquinated accumulations of mutant TDP-43 have been linked to the development of ALS and frontotemporal lobar degeneration.

The Snca^A30P allele mimics a human mutation associated with synucleinopathies such as Parkinson's disease and multiple system atrophy, as well as Lewy body dementia. The endogenous Snca gene encodes alpha-synuclein which is expressed at high levels in neurons. Mutations in human SNCA that result in aberrant polymerization and fibrillar aggregations are associated with neurodegenerative synucleinopathies.

Mapt^P290L allele encodes a microtubule-associated protein tau protein mutation that is equivalent to the human MAPT P301L mutation identified in FTDP17 (frontotemporal dementia and parkinsonism linked to chromosome 17).

Development

Stock No. 034028 Tardbp^G348C ; Snca^A30P ; Mapt^P290L(aka, Tardbp G348C KI ; Snca A30P ; Mapt V289V P290L) is a triple mutant strain assembled in the laboratory of Dr. Kristen O'Connell (The Jackson Laboratory) by breeding together multiple C57BL/6J-congenic strains and selecting for the mutant alleles listed below.

The Tardbp^G348C (Tardbp G348C KI) allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing by Dr. Cathleen Lutz (The Jackson Laboratory). Plasmids encoding a signal guide RNA were designed to introduce a glycine-to-cysteine G348C mutation in the TAR DNA binding protein locus (Tardbp) on Chromosome 4. These sequences and Cas9 nuclease were introduced into the cytoplasm C57BL/6J-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Progeny were screened by DNA sequencing of the targeted region, which identified a founder mouse harboring the desired allele. This founder was bred to C57BL/6J mice (Stock No. 000664) for germline transmission. The Tardbp^G348C colony was thereafter maintained on the C57BL/6J genetic background (see Stock No. 028435).

The Snca^A30P (Snca A30P) allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing by Dr. Cathleen Lutz (The Jackson Laboratory). Guide RNAs were selected to target exon 2 of the synuclein, alpha locus (Snca) on Chromosome 6. Donor DNAs were created encoding a alanine-to-proline mutation A30P (GCT->CCT) and a silent point mutation E28E (GAG->GAA). Ensembl transcript ID ENSMUST00000114268.4 (Snca-201) was used as reference for the exon numbering and the guide/donor sequences. These sequences and Cas9 nuclease were introduced into C57BL/6J zygotes, and then transferred to pseudopregnant females. Progeny were screened by DNA sequencing of the targeted region, which identified a founder mouse harboring the desired allele. This founder was bred to C57BL/6J mice (Stock No. 000664) for germline transmission. The Snca^A30P colony was thereafter maintained on the C57BL/6J genetic background.

The Mapt^P290L (Mapt V289V P290L) allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing by Dr. Cathleen Lutz (The Jackson Laboratory). Guide RNAs were selected to target exon 9 of the microtubule-associated protein tau locus (Mapt) on Chromosome 11. Donor DNAs were created encoding a proline-to-leucine mutation P290L (CCG->CTG) and a silent point mutation V289V (GTC->GTT). Ensembl transcript ID ENSMUST00000100347.10 (Mapt-201) was used as reference for the exon numbering and the guide/donor sequences. These sequences and Cas9 nuclease were introduced into C57BL/6J zygotes, and then transferred to pseudopregnant females. Progeny were screened by DNA sequencing of the targeted region, which identified a founder mouse harboring the desired allele. This founder was bred to C57BL/6J mice (Stock No. 000664) for germline transmission. The Mapt^P290L colony was thereafter maintained on the C57BL/6J genetic background.

Allele

Symbol: Mapt^em2Lutzy
Name: endonuclease-mediated mutation 2, Cathy Lutz
MGI accession ID: MGI:8166666
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Marker symbol: Mapt
Marker name: microtubule-associated protein tau
Marker synonyms: DDPAC; FTD1; FTDP-17; MAPT_0N4R; MAPT_1N4R; MAPT_2N4R; MAPTL; MSTD; Mtapt; MTBT1; MTBT2; PPND; PPP1R103; pTau; RNPTAU; Tau; tau-40; Tau-PHF6
Chromosome: 11
Strain of origin: C57BL/6J
Molecular note: The allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing using guide RNAs selected to target exon 9 of the microtubule-associated protein tau locus (Mapt) on Chromosome 11. Donor DNAs were created encoding a proline-to-leucine mutation P290L (CCG->CTG) and a silent point mutation V289V (GTC->GTT).

Allele

Symbol: Tardbp^em9Lutzy
Name: endonuclease-mediated mutation 9, Cathy Lutz
MGI accession ID: MGI:6295441
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: Tardbp^G348C
Marker symbol: Tardbp
Marker name: TAR DNA binding protein
Marker synonyms: 1190002A23Rik; ALS10; Tdp43; TDP-43
Chromosome: 4
Strain of origin: C57BL/6J
Molecular note: CRISPR/Cas9 genome editing is used to introduce a G348C amino acid substitution. In humans, the G348C substitution is associated with amyotrophic lateral sclerosis.

Allele

Symbol: Snca^em2Lutzy
Name: endonuclease-mediated mutation 2, Cathy Lutz
MGI accession ID: MGI:8166665
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Marker symbol: Snca
Marker name: synuclein, alpha
Marker synonyms: alphaSYN; alpha-Syn; alpha-synuclein; NACP; PARK1; PARK4; PD1
Chromosome: 6
Strain of origin: C57BL/6J
Molecular note: The allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing by using guide RNAs selected to target exon 2 of the synuclein, alpha locus (Snca) on Chromosome 6. Donor DNAs were created encoding a alanine-to-proline mutation A30P (GCT>CCT) and a silent point mutation E28E (GAG>GAA).