Luciferase-Specific Coelenterazine Analogues for Optical Cross Talk-Free Bioassays
Luciferase-Specific Coelenterazine Analogues for Optical Cross Talk-Free Bioassays
复制标题
用于无光学串扰生物测定的荧光素酶特异性腔肠素类似物
DOI：
10.1007/978-1-0716-1258-3_12
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Kim Sung-Bae
中科院分区：
文献类型：
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作者：
Nishihara Ryo;Abe Masahiro;Suzuki Koji;Kim Sung-Bae
荧光（FL）和生物发光（BL）的光谱重叠通常会导致光学串扰。本方案引入了五种不同谱系的腔肠素（CTZ）类似物，其对特定荧光素酶具有选择性，因此无串扰。例如，一些具有乙炔基或苯乙烯基的CTZ类似物通过修饰CTZ的咪唑并吡啶骨架的C-2和C-6位置处的官能团而显示出对特定酶和pH的显著偏向的BL。的光学串扰的特点是例示与多重系统，它同时照亮抗雌激素和雷帕霉素的活动，没有光学串扰。这种独特的方案有助于在没有光学污染的情况下对哺乳动物细胞中的多个光学读数进行特异性和高通量的BL成像。
Spectral overlaps in fluorescence (FL) and bioluminescence (BL) commonly cause optical cross talks. The present protocol introduces five different lineages of coelenterazine (CTZ) analogues, which have selectivity to a specific luciferase, and thus cross talk-free. For example, some CTZ analogues with ethynyl or styryl groups display dramatically biased BL to specific luciferases and pH by modifying the functional groups at the C-2 and C-6 positions of the imidazopyridinne backbone of CTZ. The optical cross talk-free feature is exemplified with the multiplex system, which simultaneously illuminated antiestrogenic and rapamycin activities without optical cross talks. This unique protocol contributes to specific and high-throughput BL imaging of multiple optical readouts in mammalian cells without optical contamination.