Patrick P Stephen, Lyons Scott K, Rodrigues Tiago B, Brindle Kevin M
Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, UK, CB2 1QW,
Mol Imaging Biol. 2014 Oct;16(5):626-34. doi: 10.1007/s11307-014-0741-4.
Bioluminescence imaging is a powerful tool for studying gene expression and cell migration in intact living organisms. However, production of bioluminescence by cells transfected to express luciferase can be limited by the rate of plasma membrane transport of its substrate D-luciferin. We sought to identify a plasma membrane transporter for D-luciferin that could be expressed alongside luciferase to increase transmembrane flux of its substrate and thereby increase light output.
Luciferase-expressing cells were transfected with a lentivirus encoding the rat reno-hepatic organic anion transporter protein, Oatp1, which was identified as a potential transporter for D-luciferin. Light output was compared between cells expressing luciferase and those also expressing Oatp1.
In two cell lines and in mouse xenografts, co-expression of Oatp1 with luciferase increased light output by several fold, following addition of luciferin.
The increase in light output thus obtained will allow more sensitive detection of luciferase-expressing cells in vivo.
生物发光成像技术是研究完整活生物体中基因表达和细胞迁移的有力工具。然而，转染表达荧光素酶的细胞产生的生物发光可能会受到其底物D - 荧光素质膜转运速率的限制。我们试图鉴定一种D - 荧光素的质膜转运体，它可以与荧光素酶一起表达，以增加其底物的跨膜通量，从而增加光输出。
用编码大鼠肾肝有机阴离子转运蛋白Oatp1的慢病毒转染表达荧光素酶的细胞，该蛋白被鉴定为D - 荧光素的潜在转运体。比较了表达荧光素酶的细胞与同时表达Oatp1的细胞之间的光输出。
在两种细胞系和小鼠异种移植模型中，加入荧光素后，Oatp1与荧光素酶的共表达使光输出增加了几倍。
由此获得的光输出增加将使体内表达荧光素酶的细胞检测更加灵敏。
