Vývoj a využití deoxyribozymů, které produkují barvu a fluorescenci

Volek, Martin

1 Abstract The recent SARS-CoV-2 pandemic showed how humanity can be threatened by organisms as simple as viruses. This crisis emphasized the need for timely and reliable diagnostics, as well as the fast discovery of effective therapeutics against new diseases. We propose that both challenges can be addressed by deoxyribozymes (catalytically active DNA molecules). The most common types of signals are light, fluorescence, color and radioactivity. Some of these signals can be generated by deoxyribozymes, but others cannot. A functional DNA molecule that generates a robust fluorogenic or chromogenic signal would have significant advantages for many applications compared to intrinsically fluorescent proteins, which are expensive and labour intensive to synthesize, and fluorescent RNA aptamers, which are more prone to degradation than DNA molecules. To broaden the spectrum of possible outcomes generated by DNA, we have used in vitro selection to develop deoxyribozymes, which generate strong fluorogenic and chromogenic signals. Deoxyribozymes that generate a fluorogenic signal achieve this by dephosphorylating the coumarin substrate 4-methylumbelliferyl phosphate (4-MUP). They transfer the phosphate group from 4-MUP to their own 5' hydroxyl group. This reaction converts the non-fluorogenic 4-MUP into the...

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