Aurora: a fluorescent deoxyribozyme for high-throughput screening

被引:3
作者
Volek, Martin [1 ,2 ]
Kurfurst, Jaroslav [1 ,3 ]
Drexler, Matus [1 ]
Svoboda, Michal [1 ]
Srb, Pavel [1 ]
Veverka, Vaclav [1 ,4 ]
Curtis, Edward A. [1 ]
机构
[1] Czech Acad Sci, Inst Organ Chem & Biochem, Prague 16610, Czech Republic
[2] Charles Univ Prague, Fac Sci, Dept Genet & Microbiol, Prague 12844, Czech Republic
[3] Univ Chem & Technol, Dept Informat & Chem, Prague 16628, Czech Republic
[4] Charles Univ Prague, Fac Sci, Dept Cell Biol, Prague 12844, Czech Republic
关键词
IN-VITRO SELECTION; DNA APTAMER PAIR; ALLOSTERIC RIBOZYME; RNA; CHEMILUMINESCENCE; FLUOROPHORE; GENERATION; MOLECULES; MECHANISM; PROTEIN;
D O I
10.1093/nar/gkae467
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Fluorescence facilitates the detection, visualization, and tracking of molecules with high sensitivity and specificity. A functional DNA molecule that generates a robust fluorescent signal would offer significant advantages for many applications compared to intrinsically fluorescent proteins, which are expensive and labor intensive to synthesize, and fluorescent RNA aptamers, which are unstable under most conditions. Here, we describe a novel deoxyriboyzme that rapidly and efficiently generates a stable fluorescent product using a readily available coumarin substrate. An engineered version can detect picomolar concentrations of ribonucleases in a simple homogenous assay, and was used to rapidly identify novel inhibitors of the SARS-CoV-2 ribonuclease Nsp15 in a high-throughput screen. Our work adds an important new component to the toolkit of functional DNA parts, and also demonstrates how catalytic DNA motifs can be used to solve real-world problems. Graphical Abstract
引用
收藏
页码:9049 / 9061
页数:13
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