title: The Terminator mouse is a diphtheria toxin-receptor knock-in mouse strain for rapid and efficient enrichment of desired cell lineages.
abstract: Biomedical research often requires primary cultures of specific cell types, which are challenging to obtain at high purity in a reproducible manner. Here we engineered the murine Rosa26 locus by introducing the diphtheria toxin receptor flanked by loxP sites. The resultant strain was nicknamed the Terminator mouse. This approach results in diphtheria toxin-receptor expression in all non-Cre expressing cell types, making these cells susceptible to diphtheria toxin exposure. In primary cultures of kidney cells derived from the Terminator mouse, over 99.99% of cells were dead within 72 h of diphtheria toxin treatment. After crossing the Terminator with the podocin-Cre (podocyte specific) mouse or the Ggt-Cre (proximal tubule specific) mouse, diphtheria toxin treatment killed non-Cre expressing cells but spared podocytes and proximal tubule cells, respectively, enriching the primary cultures to over 99% purity, based on both western blotting and immunostaining of marker proteins. Thus, the Terminator mouse can be a useful tool to selectively and reproducibly obtain even low-abundant cell types at high quantity and purity.
authors.affiliation: Department of Internal Medicine/Section of Nephrology, Yale University School of Medicine, New Haven, Connecticut, USA.
authors.forename: Jian-Kan
authors.initials: JK
authors.lastname: Guo
authors.forename: Hongmei
authors.initials: H
authors.lastname: Shi
authors.forename: Farrukh
authors.initials: F
authors.lastname: Koraishy
authors.forename: Arnaud
authors.initials: A
authors.lastname: Marlier
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Ding
authors.forename: Alan
authors.initials: A
authors.lastname: Shan
authors.forename: Lloyd G
authors.initials: LG
authors.lastname: Cantley
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D019070:Cell Lineage; D002469:Cell Separation; D002470:Cell Survival; D004167:Diphtheria Toxin / Q000494:pharmacology; D005838:Genotype; D066257:Heparin-binding EGF-like Growth Factor; D007150:Immunohistochemistry; D019426:Integrases / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010641:Phenotype; D050199:Podocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D061251:Primary Cell Culture; D022661:RNA, Untranslated / Q000235:genetics; D013997:Time Factors
chemical_list: D015415:Biomarkers; D004167:Diphtheria Toxin; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D022661:RNA, Untranslated; C045073:Cre recombinase; D019426:Integrases