B6(Cg)-Zbtb46^{tm1(HBEGF)Mnz}/J

Stock number: 019506
Product name: zDC-DTR
Research areas: Research Tools

Description

A cDNA encoding human diptheria toxin receptor (DTR; formal symbol HBEGF) was introduced into the 3' untranslated region (UTR) of the mouse Zbtb46 (zinc finger and BTB domain containing 46; also called zDC) gene. Unlike previous CD11c-DTR models, classical dendritic cells (cDCs) can be specifically depleted from bone marrow chimeras through diptheria toxin (DT) treatment. Non-cDCs (including pDCs, monocytes, macrophages and NK cells) are not affected.

Detailed description

Zbtb46 (zinc finger and BTB domain containing 46; also called zDC) is expressed by classical dendritic cells (cDCs) and committed cDC precursors but not monocytes or other immune cell populations. As such, it is a marker that defines the classical dendritic cell lineage, but it is also expressed by an undefined radioresistant, non-hematopoietic cell population.

A cDNA encoding human diptheria toxin receptor (DTR; formal symbol HBEGF) and mCherry were introduced into the 3' untranslated region (UTR) of the mouse Zbtb46 gene. ZBTB46 protein is expressed at physiological levels from the knockin allele. This mutation serves as an indicator of gene expression and creates a means to specifically deplete cDCs. Although DTR is detectable by flow cytometry, mCherry fluorescence cannot be directly detected.

Injection of a single dose of diptheria toxin (DT) in knockin mice and C57BL/6-to-knockin bone marrow chimeras is fatal within 24-48 hours. DT treatment of knockin-to-C57BL/6 bone marrow chimeras, however can reportedly survive DT injections every other day for more than 2 weeks. Unlike previous CD11c-DTR models, cDCs are specifically depleted, but non-cDCs (including pDCs, monocytes, macrophages and NK cells) are not affected by toxin treatment.

Development

A targeting vector containing an ACN self-excising loxP-flanked neomycin cassette was used to introduce IRES, a cDNA encoding human diptheria toxin receptor (DTR) and mCherry immediately after the endogenous stop codon in exon 5. The mutation was created in CY2.4 B6(Cg)-Tyr^c-2J-derived ES cells. Chimeric male progeny were crossed with B6(Cg)-Tyr^c-2J females to allow for coat color screening of germline transmission and for self-excision of the ACN cassette. White animals were crossed to C57BL/6 for four generations by the donating laboratory, and both white and black animals were retained in their colony.

Allele

Symbol: Zbtb46^{tm1(HBEGF)Mnz}
Name: targeted mutation 1, Michel C Nussenzweig
MGI accession ID: MGI:5444842
Generation method: Targeted
Attributes: Reporter; Inserted expressed sequence
Synonyms: Zbtb46^DTR; Zbtb46^tm1(DTR)Mnz; zDC-DTR
Marker symbol: Zbtb46
Marker name: zinc finger and BTB domain containing 46
Marker synonyms: 2610019F01Rik; 4933406L05Rik; Btbd4; BZEL; dJ583P15.7; dJ583P15.8; RINZF; ZNF340
Chromosome: 2
Strain of origin: B6(Cg)-Tyr^c-2J
Expressed genes: HBEGF,heparin binding EGF like growth factor,human; DTR,Simian Diphtheria Toxin Receptor,
Molecular note: An IRES and human diphtheria toxin receptor (HBEGF) fused with mCherry was inserted downstream of the stop codon. Flow cytometry failed to detect expression of mCherry.