Molecular Engineering of Thiazole Orange Dye: Change of Fluorescent Signaling from Universal to Specific upon Binding with Nucleic Acids in Bioassay
Molecular Engineering of Thiazole Orange Dye: Change of Fluorescent Signaling from Universal to Specific upon Binding with Nucleic Acids in Bioassay
复制标题
噻唑橙染料的分子工程：生物测定中与核酸结合后荧光信号从通用到特异性的变化
DOI：
10.1021/acschembio.5b00987
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发表时间：
2016-04-01
影响因子：
4
通讯作者：
Chow, Cheuk-Fai
中科院分区：
文献类型：
--
作者：
Lu, Yu-Jing;Deng, Qiang;Chow, Cheuk-Fai
噻唑橙子（TO）染料的通用荧光染色性质通过在TO支架的邻位引入苯乙烯样取代基而被调整，以便对G-四链体DNA结构具有特异性。这一非凡的结果是从实验研究中确定的，并通过分子对接研究进一步探索。分子对接研究有助于理解这样一个小的取代基如何导致G-四链体DNA和其他类型的核酸之间显着的荧光信号区分。结果表明，修饰的染料以与TO类似的方式结合到G-四链体或双链体DNA上，但表现出增强或淬灭的荧光信号，这是由取代基的空间长度和取向决定的，并且从未被发现过。新的荧光染料与p-（二甲基氨基）苯乙烯基取代基修饰提供了10倍以上的选择性端粒G-四链体比双链DNA基板。此外，还研究了非变性PAGE实验、FRET、CD分析和活细胞成像，并证明了这类基于噻唑橙的荧光探针在生物测定和细胞成像中的潜在应用。
The universal fluorescent staining property of thiazole orange (TO) dye was adapted in order to be specific for G-quadruplex DNA structures, through the introduction of a styrene-like substituent at the ortho-position of the TO scaffold. This extraordinary outcome was determined from experimental studies and further explored through molecular docking studies. The molecular docking studies help understand how such a small substituent leads to remarkable fluorescent signal discrimination between G-quadruplex DNA and other types of nucleic acids. The results reveal that the modified dyes bind to the G-quadruplex or duplex DNA in a similar fashion as TO, but exhibit either enhanced or quenched fluorescent signal, which is determined by the spatial length and orientation of the substituent and has never been known. The new fluorescent dye modified with a p-(dimethylamino)styryl substituent offers 10-fold more selectivity toward telomeric G-quadruplexes than double-stranded DNA substrates. In addition, native PAGE experiments, FRET, CD analysis, and live cell imaging were also studied and demonstrated the potential applications of this class of thiazole-orange-based fluorescent probes in bioassays and cell imaging.