The first 6 exons of Ebf3 were replaced with a cassette encoding OR8A1 (formerly OLFR151) followed by an IRES and Tau-GFP for bicistronic expression driven by the Ebf3 promoter. This gives robust expression of Or8a1 mRNA and widespread GFP fluorescence in the main olfactory epithelium and vomeronasal organ in heterozygotes. In general, the glomeruli have normal morphology and the main olfactory epithelium has a normal multi-layered structure and thickness even though the OR8A1 expression is found at the immature GAP43- stage of olfactory sensory neuron development. Although olfactory sensory neurons usually express only one olfactory receptor gene and exclude expression off all others, these OR8A1+ neurons co-express a second class II olfactory receptor gene from the normal, endogenous expression process. Nanostring RNA analysis of nineteen olfactory receptor genes found normal expression levels of eighteen and a slight reduction in expression level of only one. Patch-clamp assessment of these GFP+ olfactory sensory neurons expressing ectopic OLFR151 respond to acetophenone, a known ligand for OR8A1.

Targeted replacement of the first six exons of Ebf3 with a cassette encoding OR8A1-IRES-Tau-GFP gives heterozygotes widespread bicistronic expression of OR8A1 (formerly OLFR151) and Tau-GFP throughout the main olfactory epithelium and vomeronasal organ but does not prevent endogenous olfactory receptor expression. Thus, more than one olfactory receptor protein is expressed on these olfactory sensory neurons.

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