B6.Cg-Apoe^{em3(APOE*)Adiuj}/J

Stock number: 029017
Product name: APOE2 KI
Strain synonyms: APOE*2 KI
Research areas: Cardiovascular Research, Metabolism Research, Neurobiology Research, Research Tools

Description

APOE*2 KI are CRISPR/Cas9-generated mice with R130C and R176C point mutations in the human APOE sequence of an existing humanized ApoE knock-in allele and express the human APOE isoform E2.

Detailed description

The targeted Apoe gene encodes apolipoprotein E, which is important in lipoprotein metabolism and cardiovascular disease as well as Alzheimer's disease, immunoregulation and cognition. CRISPR/Cas9 was used to introduce a C to T nucleotide substitution point mutation in amino acids R130C and R176C into the human APOE sequence in the Apoe^{tm1.1(APOE*4)Adiuj} allele. The resulting APOE*2 KI mice express APOE isoform 2, from the endogenous Apoe locus. The APOE 2 isoform is found in approximately 7% of the population, and does not bind to cell surface receptors as well as the 3 and 4 isoforms. As the mice are characterized, we may modify the strain description accordingly.

The Jackson Laboratory colony shows that homozygous mice of both sexes are viable and fertile (April 2021).

Development

A mutagenic oligo based approach was used to generate this strain. Plasmids encoding a signal guide RNA designed to introduce the R130C and R176C point mutations into the Apoe^{tm1.1(APOE*4)Adiuj} allele and the cas9 nuclease were introduced into the cytoplasm of B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj}/J (Stock No. 027894)-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.

Allele

Symbol: Apoe^{em3(APOE*)Adiuj}
Name: endonuclease-mediated mutation 3, MODEL-AD Center
MGI accession ID: MGI:6164017
Generation method: Endonuclease-mediated
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Apoe
Marker name: apolipoprotein E
Marker synonyms: AD2; APO-E; ApoE4; APOEA; LDLCQ5; LPG
Chromosome: 7
Strain of origin: B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj}/J
Expressed genes: APOE,apolipoprotein E,human
Molecular note: CRISPR/Cas9 genome editing methodologies was used to introduce p.R130C and p.R176C point mutations into human APOE protein carried by the Apoe^{tm1.1(APOE*4)Adpmc} allele.