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However, their isoform-specific detection remains challenging. To facilitate the analysis of G\u03b1<jats:sub>i3<\/jats:sub>expression, we generated a<jats:italic>Gnai3\ufeff\ufeff-<\/jats:italic>\ufeffiresGFP reporter mouse line. An internal ribosomal entry site (IRES) was inserted behind the stop-codon of the<jats:italic>Gnai3<\/jats:italic>gene to initiate simultaneous translation of the GFP cDNA together with G\u03b1<jats:sub>i3<\/jats:sub>. The expression of GFP was confirmed in spleen and thymus tissue by immunoblot analysis. Importantly, the GFP knock-in (ki) did not alter G\u03b1<jats:sub>i3<\/jats:sub>expression levels in all organs tested including spleen and thymus compared to wild-type littermates. Flow cytometry of thymocytes, splenic and blood cell suspensions revealed significantly higher GFP fluorescence intensities in homozygous ki\/ki animals compared to heterozygous mice (+\/ki). Using cell-type specific surface markers GFP fluorescence was assigned to B cells, T cells, macrophages and granulocytes from both splenic and blood cells and additionally blood-derived platelets. Moreover, immunofluorescent staining of the inner ear from knock-in mice unraveled GFP expression in sensory and non-sensory cell types, with highest levels in Deiter\u2019s cells and in the first row of Hensen\u2019s cells in the organ of Corti, indicating a novel site for G\u03b1<jats:sub>i3<\/jats:sub>expression. In summary, the<jats:italic>Gnai3-\ufeff\ufeff<\/jats:italic>iresGFP\u00a0reporter mouse represents an ideal tool for precise analyses of G\u03b1<jats:sub>i3<\/jats:sub>expression patterns and sites.<\/jats:p>","DOI":"10.1038\/s41598-021-93591-0","type":"journal-article","created":{"date-parts":[[2021,7,12]],"date-time":"2021-07-12T10:03:06Z","timestamp":1626084186000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Analyses of Gnai3-iresGFP reporter mice reveal unknown G\u03b1i3 expression sites"],"prefix":"10.1038","volume":"11","author":[{"given":"Veronika","family":"Leiss","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ellen","family":"Reisinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annika","family":"Speidel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandra","family":"Beer-Hammer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernd","family":"N\u00fcrnberg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,7,12]]},"reference":[{"key":"93591_CR1","doi-asserted-by":"publisher","first-page":"615","DOI":"10.1146\/annurev.bi.56.070187.003151","volume":"56","author":"AG Gilman","year":"1987","unstructured":"Gilman, A. 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