Using CRISPR gene editing technology, a V5-Avi-IRES-Akaluc-2A-tdTomato expression cassette was inserted before the stop codon TGA of mouse Foxp3 gene to establish a multi-labeled Knock-in model with V5, Avi, Akaluc luciferase and tdTomato fluorescent protein. Akaluc is a novel luciferase that produces up to 1,000 times as bright as light emitted by existing systems by feeding or injecting a synthetic luciferin compound called AkaLumine-HCl, which is better able to penetrate the blood-brain barrier than d-luciferin is.
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