Firefly Luciferase Mutants Allow Substrate-Selective Bioluminescence Imaging in the Mouse Brain.
Firefly Luciferase Mutants Allow Substrate-Selective Bioluminescence Imaging in the Mouse Brain.
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DOI：
10.1002/anie.201511350
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发表时间：
2016-04-11
期刊：
影响因子：
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通讯作者：
Miller SC
中科院分区：
文献类型：
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作者：
Adams ST Jr;Mofford DM;Reddy GS;Miller SC
生物发光成像是一种强大的方法来可视化活体小鼠体内发生的特定事件。可以使动物对基因的表达、酶的活性或肿瘤的生长做出反应而发光。但生物发光需要荧光素酶与小分子荧光素的相互作用，其范围受到少数自然组合的限制。在这里，我们表明，萤火虫荧光素酶的突变体可以区分天然和合成底物在活小鼠的大脑。使用腺相关病毒（AAV）载体在大脑中表达腺苷酸酶，我们发现，突变体腺苷酸酶是无活性或弱活性的D-腺苷酸可以明亮地亮起时，与氨基荧光素CycLuc 1和CycLuc 2或其各自的FAAH敏感的腺苷酸酰胺处理。选择性荧光素酶的进一步发展有望扩大生物发光的能力，并允许在同一活体动物中对多个事件进行成像。
Bioluminescence imaging is a powerful approach to visualize specific events occurring inside live mice. Animals can be made to glow in response to the expression of a gene, the activity of an enzyme, or the growth of a tumor. But bioluminescence requires the interaction of a luciferase enzyme with a small molecule luciferin, and its scope has been limited by the mere handful of natural combinations. Here we show that mutants of firefly luciferase can discriminate between natural and synthetic substrates in the brains of live mice. Using adeno-associated viral (AAV) vectors to express luciferases in the brain, we find that mutant luciferases that are inactive or weakly active with D-luciferin can light up brightly when treated with the aminoluciferins CycLuc1 and CycLuc2 or their respective FAAH-sensitive luciferin amides. Further development of selective luciferases promises to expand the power of bioluminescence and allow multiple events to be imaged in the same live animal.