NOD.Cg-Mertk^em1Doi Prkdc^scid/RweiJ

Stock number: 036071
Product name: Mertk^GFPiDTR , Mertk-GFP-DTR
Research areas: Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

Mertk-GFP-DTR knock-in/knock-out mice express the EGFP reporter and the diphtheria toxin receptor (DTR) under the control of the Mertk (MER proto-oncogene tyrosine kinase) promoter. GFP expression is restricted to macrophages. This strain may be useful in tracing and ablating macrophages.

Detailed description

NOD SCID Mertk-GFP-DTR mutant mice contain an EGFP reporter and cDNA encoding human diptheria toxin receptor (DTR; formal symbol HBEGF) inserted into the translational start site of the Mertk (MER proto-oncogene tyrosine kinase) gene. Mertk encodes a transmembrane protein that functions as a receptor kinase and is expressed in macrophages. Mice homozygous for scid and heterozygous for Mertk-GFP-DTR are viable and fertile. Homozygosity for Mertk-GFP-DTR results in poor health above 25 weeks of age. GFP fluorescence is restricted to macrophages and exhibits some heterogeneity especially in islet-resident macrophages. NOD SCID Mertk-GFP-DTR mice do not develop spontaneous diabetes. This strain may be useful in tracing and ablating macrophages.

Development

The Mertk^em1Doi allele was created in the laboratory of Dr. Diane Mathis at Harvard Medical School. CRISPR/Cas9 endonuclease-mediated genome editing was used to insert a construct containing an EGFP, an IRES, a human diphtheria toxin receptor (DTR) followed by a polyA tail and a 3' HA tag after the ATG site in exon 1 of the Mertk (MER proto-oncogene tyrosine kinase) gene on chromosome 2. The insertion results in a 53 bp deletion in the remainder of exon 1. Single guide RNA and Cas9 endonuclease were introduced into NOD-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to NOD mice for germline transmission. Dr. Ralph Weissleder (Massachusetts General Hospital) obtained Mertk-GFP-DTR from Dr. Mathis and introgressed the allele into the NOD.Cg-Prkdc^scid background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize NOD.Cg-Prkdc^scid/J (Stock No. 001303) oocytes.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Synonyms: scid
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Marker synonyms: DNAPDcs; DNAPK; DNA-PK; DNAPKc; DNA-PKC; DNA-PKcs; DNPK1; DOXNPH; dxnph; HYRC; HYRC1; IMD26; p350; S473K; slip; XRCC7
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).

Allele

Symbol: Mertk^em1Doi
Name: endonuclease-mediated mutation 1, Christophe Benoist and Diane Mathis
MGI accession ID: MGI:6386251
Generation method: Endonuclease-mediated
Attributes: Reporter; Epitope tag
Synonyms: Mertk^GFPiDTR
Marker symbol: Mertk
Marker name: c-mer proto-oncogene tyrosine kinase
Marker synonyms: c-Eyk; c-mer; Eyk; MER; nmf12; Nyk; rdy; RP38; Tyro 12; Tyro12
Chromosome: 2
Strain of origin: NOD
Molecular note: CRISPR/Cas9 genome editing is used to introduce a 5' HA tag, an EGFP reporter, an IRES, and a DTR with an SV40 polyadenylation sequence and 3' HA tag into exon 1, following the ATG codon. The insertion results in a 53 bp deletion in the remainder of exon 1.