Homologous recombination into the eosinophil peroxidase locus generates a strain of mice expressing Cre recombinase exclusively in eosinophils

Cre‐recombinase expression in eoCRE mice generates a model system facilitating eosinophil‐specific knockouts, and/or, ectopic overexpression of heterologous genes. Eosinophils are generally linked to innate host defense against helminths, as well as the pathologies associated with allergic diseases,...

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Published in:	Journal of leukocyte biology Vol. 94; no. 1; pp. 17 - 24
Main Authors:	Doyle, Alfred D., Jacobsen, Elizabeth A., Ochkur, Sergei I., Willetts, Lian, Shim, Kelly, Neely, Joseph, Kloeber, Jake, LeSuer, Will E., Pero, Ralph S., Lacy, Paige, Moqbel, Redwan, Lee, Nancy A., Lee, James J.
Format:	Journal Article
Language:	English
Published:	United States Society for Leukocyte Biology 01-07-2013
Subjects:	Animals Bone Marrow - metabolism Cell Differentiation Cell Lineage Cell Proliferation Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism eosinophil granule protein eosinophil lineage‐specific knock‐out Eosinophil Peroxidase - physiology Eosinophils - enzymology EPX Flow Cytometry Homologous Recombination Humans Integrases - metabolism knock‐in mice Mice Mice, Inbred C57BL Mice, Transgenic Spotlight on Leading Edge Research eosinophil granule protein knock-in mice eosinophil lineage-specific knock-out EPX
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Summary:	Cre‐recombinase expression in eoCRE mice generates a model system facilitating eosinophil‐specific knockouts, and/or, ectopic overexpression of heterologous genes. Eosinophils are generally linked to innate host defense against helminths, as well as the pathologies associated with allergic diseases, such as asthma. Nonetheless, the activities of eosinophils remain poorly understood, which in turn, has prevented detailed definitions of their role(s) in health and disease. Homologous recombination in embryonic stem cells was used to insert a mammalianized Cre recombinase in the ORF encoding Epx. This knock‐in strategy overcame previous inefficiencies associated with eosinophil‐specific transgenic approaches and led to the development of a knock‐in strain of mice (eoCRE), capable of mediating recombination of “floxed” reporter cassettes in >95% of peripheral blood eosinophils. We also showed that this Cre expression was limited exclusively to eosinophil‐lineage committed cells with no evidence of Cre‐mediated toxicity. The efficiency and specificity of Cre expression in eoCRE mice were demonstrated further in a cross with a knock‐in mouse containing a “(flox‐stop‐flox)” DTA cassette at the ROSA26 locus, generating yet another novel, eosinophil‐less strain of mice. The development of eoCRE mice represents a milestone in studies of eosinophil biology, permitting eosinophil‐specific gene targeting and overexpression in the mouse as part of next‐generation studies attempting to define eosinophil effector functions.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	0741-5400 1938-3673
DOI:	10.1189/jlb.0213089