Enhanced synthetic luciferin substrates for firefly luciferase

生物发光分析和成像方法已被广泛应用于 药物发现，因为它们的简单性、健壮性和低成本。与生俱来的低 背景和不需要激发光使生物发光优于 荧光方法有多种应用。尽管如此，生物发光还没有 充分发挥其潜力。我们提出以下具体目标：1)扩大 利用人工合成的荧光素进行生物发光成像；2)发展明亮和功能 适用于活体成像的正交型荧光素酶；3)利用“便携式记者” 在任何动物身上进行成像。我们将利用合成的荧光素类似物的特性来 增强侵袭性脑肿瘤、中枢神经系统病毒感染和昼夜节律的成像 节奏。我们将开发完全正交的荧光素-荧光素酶对，可以作为 内部控制，并启用多路复用成像。最后，我们将克服限制 通过开发生物发光记者获得一组可用的表达荧光素酶的动物 可以被引入到任何感兴趣的动物中。

Bioluminescence assays and imaging methods have been widely adopted for drug discovery because of their simplicity, robustness, and low-cost. The inherently low background and lack of need for excitation light makes bioluminescence superior to fluorescence methods for many applications. Nevertheless, bioluminescence has yet to reach its full potential. We propose the following specific aims: 1) Expand the scope of bioluminescence imaging with synthetic luciferins; 2) Develop bright and functionally orthogonal luciferases suitable for in vivo imaging; and 3) Leverage “portable reporters” for imaging in any animal. We will exploit the properties of synthetic luciferin analogs to enhance the imaging of invasive brain tumors, viral infection of the CNS, and circadian rhythms. We will develop fully orthogonal luciferin-luciferase pairs that can serve as internal controls and enable multiplexed imaging. Finally, we will overcome the restricted set of available luciferase-expressing animals by developing bioluminescent reporters that can be introduced into any animal of interest.