Moroz Maxim A, Zurita Juan, Moroz Anna, Nikolov Ekaterina, Likar Yury, Dobrenkov Konstantin, Lee Jason, Shenker Larissa, Blasberg Ronald, Serganova Inna, Ponomarev Vladimir
Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
Skolkovo Institute of Science and Technology, Moscow 143026, Russia.
Mol Ther Oncolytics. 2021 Mar 17;21:15-22. doi: 10.1016/j.omto.2021.03.004. eCollection 2021 Jun 25.
Bioluminescence reporter gene imaging is a robust, high-throughput imaging modality that is useful for tracking cells and monitoring biological processes, both in cell culture and in small animals. We introduced and characterized a novel bioluminescence reporter-membrane-anchored luciferase (maCLuc)-paired with a unique vargulin substrate. This luciferase-substrate pair has no cross-reactivity with established d-luciferin- or coelenterazine-based luciferase reporters. We compare maCLuc with several established luciferase-based reporter systems (firefly, click beetle, , and luciferases), using both and models. We demonstrate the different imaging characteristics of these reporter systems, which allow for multiplexed-luciferase imaging of 3 and 4 separate targets concurrently in the same animal within 24 h. The imaging paradigms described here can be directly applied for simultaneous monitoring of multiple cell populations, the activity of selected signal transduction pathways, or a combination of both constitutive and inducible reporter imaging.
生物发光报告基因成像技术是一种强大的高通量成像方式，可用于在细胞培养和小动物体内追踪细胞并监测生物过程。我们引入并表征了一种新型生物发光报告基因——膜锚定荧光素酶（maCLuc），它与一种独特的视黄醛底物配对。这种荧光素酶-底物对与已有的基于d-荧光素或腔肠素的荧光素酶报告基因没有交叉反应。我们使用细胞和动物模型，将maCLuc与几种已有的基于荧光素酶的报告系统（萤火虫荧光素酶、叩头虫荧光素酶等）进行比较。我们展示了这些报告系统不同的成像特性，这使得在24小时内能够在同一只动物体内同时对3个和4个独立目标进行多重荧光素酶成像。这里描述的成像模式可直接应用于同时监测多个细胞群体、所选信号转导途径的活性，或组成型和诱导型报告基因成像的组合。
