B6(SJL)-Tardbp^em9Lutzy Apoe^{tm1.1(APOE*4)Adiuj} Mapt^em2Lutzy/KmsoJ

Stock number: 034545
Product name: Tardbp^G348C ; APOE4 KI ; Mapt^P290L
Strain synonyms: Tardbp G348C KI ; APOE*4 KI ; Mapt V289V P290L
Research areas: Cardiovascular Research, Metabolism Research, Mouse/Human Gene Homologs, Neurobiology Research, Research Tools

Description

The Tardbp^G348C ; APOE4 KI ; 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.
APOE4 KI is a humanized ApoE knock-in mutation encoding isoform E4 suitable for use in studying lipoproteins, arteriosclerosis and coronary heart disease.
Mapt^P290L is equivalent to the human MAPT P301L mutation identified in frontotemporal dementia with parkinsonism.

Detailed description

Stock No. 034545 Tardbp^G348C ; APOE4 KI ; Mapt^P290L (aka, Tardbp G348C KI ; APOE*4 KI ; 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 APOE4 KI humanized knock-in allele has mouse Apoe exons 2, 3 and most of exon 4 replaced by a human APOE4 gene sequence including exons 2, 3 and 4 (and some 3' UTR sequence). This knock-in allele encodes apolipoprotein isoform E4, which is important in lipoprotein metabolism and cardiovascular disease as well as Alzheimer's disease, immunoregulation and cognition.

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. 034545 Tardbp^G348C ; APOE4 KI ; Mapt^P290L (aka, Tardbp G348C KI ; APOE*4 KI ; 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 APOE4 KI (APOE*4 KI) allele was generated using homologous targeting by Dr. Mike Sasner (The Jackson Laboratory) and MODEL-AD. A targeting vector containing a 3' FRT-flanked PGK-neo cassette was used to replace the mouse apolipoprotein E (Apoe) exons 2, 3 and most of exon 4 with 1.5 kb of human APOE gene sequence including exons 2, 3 and 4 (and some 3' UTR sequence). The human ApoE sequence also contains a 2 nucleotide A->T substitution in intron 3 for identification of the targeted allele by Southern analysis. The construct was electroporated into C57BL/6J derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission by crossing to C57BL/6J. Heterozygous animals were crossed subsequently to C57BL/6J-congenic FLP deleter mice (Stock No. 005703) to remove the FRT-flanked PGK-neo cassette. Mice that no longer contained the FRT-flanked PGK-neo cassette were then backcrossed to C57BL/6J at least once to remove the FLP recombinase transgene. The APOE4 KI colony was thereafter maintained on the C57BL/6J genetic background (see Stock No. 027894).

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: Apoe^{tm1.1(APOE*4)Adiuj}
Name: targeted mutation 1.1, MODEL-AD Center
MGI accession ID: MGI:5810209
Generation method: Targeted
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: APOE^epsilon4
Marker symbol: Apoe
Marker name: apolipoprotein E
Marker synonyms: AD2; APO-E; ApoE4; APOEA; LDLCQ5; LPG
Chromosome: 7
Strain of origin: C57BL/6J
Expressed genes: APOE,apolipoprotein E,human
Site of expression: Endogenous mouse elements direct human APOE gene sequence (including exons 2, 3 and 4 and some 3' UTR sequence).
Molecular note: The targeting vector contains a 3' FRT flanked neo cassette and 1.5 kb of human APOE gene sequence including exons 2, 3, 4 and a portion of the 3'UTR sequence (the APOE4 isoform) which is used to replace exons 2, 3 and most of exon 4 of the endogenous sequence. The human APOE sequence carries a 2 nucleotide A-to-T substitution in intron 3. Flp-mediated recombination removed the FRT-flanked neo cassette.