Orthogonal Luciferase–Luciferin Pairs for Bioluminescence Imaging

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• 作者：Krysten A. Jones ; William B. Porterfield ; Colin M. Rathbun ; David C. McCutcheon ; Miranda A. Paley ; Jennifer A. Prescher
• 刊名：Journal of the American Chemical Society
• 出版年：2017
• 出版时间：February 15, 2017
• 年：2017
• 卷：139
• 期：6
• 页码：2351-2358
• 全文大小：570K
• ISSN：1520-5126

文摘

Bioluminescence imaging with luciferase–luciferin pairs is widely used in biomedical research. Several luciferases have been identified in nature, and many have been adapted for tracking cells in whole animals. Unfortunately, the optimal luciferases for imaging in vivo utilize the same substrate and therefore cannot easily differentiate multiple cell types in a single subject. To develop a broader set of distinguishable probes, we crafted custom luciferins that can be selectively processed by engineered luciferases. Libraries of mutant enzymes were iteratively screened with sterically modified luciferins, and orthogonal enzyme–substrate “hits” were identified. These tools produced light when complementary enzyme–substrate partners interacted both in vitro and in cultured cell models. Based on their selectivity, these designer pairs will bolster multicomponent imaging and enable the direct interrogation of cell networks not currently possible with existing tools. Our screening platform is also general and will expedite the identification of more unique luciferases and luciferins, further expanding the bioluminescence toolkit.