Live Tracking Of The Transplanted Neural Cells By Using AkaLumine/Akaluc System In Mouse

Mechanism of CNS diseases(central nervous system disease)is complex.And neurons is out of mitosis,the number and expansion ability of adult neural stem cells is limited.So it is effect-less relying on self-repair.Cell therapy is the most hopeful way to cure nervous system disease.iNSC transplantation showed significant effect to the symptom recovery;eliminate inflammation around damaged areas and endogenous neuron degeneration.However,studies of NSC transplantation suggested that mature process of grafted NSCs would need a quite long time,from 6 months to 1 year,even more longer.During this process,how to track the status of transplanted NSCs is the key question.At present,live tracking had mainly been achieved through fluorescence and bioluminescence.While,because of the thickness of brain tissue,the absorbing light property of blood and lymphatic tissue,the ordinary fluorescence technology could not meet the needs of researches.So it is necessary to establish a more accurate system.AKalumine/AKaluc is a newly reported system based on Luciferase/Luciferin,with modified structure of luciferin and enzyme.AKalumine/AKaluc has the maximum light absorption peak near infrared reign,so it has the very strong organization transmissibility.This system has obvious advantages,such as operability and nontoxicity.In this study,we chose AAVS1,the the safety site in human genome,used CRISPR/Cas9 technology,established a AKaluc knock-in embryonic stem cell line.Then we proved the AKalumine/AKaluc could work normally in vitro and in vivo.At last,we also studied the effect of different promoteres on the expression of AKaluc,proved promoter EF1α was the better one in human ESCs.