Detection of SARS-CoV-2 RNA Using a DNA Aptamer Mimic of Green Fluorescent Protein.
ACS Chem Biol
; 17(4): 840-853, 2022 04 15.
Article
in English
| MEDLINE | ID: covidwho-1852373
ABSTRACT
RNA detection is important in diverse diagnostic and analytical applications. RNAs can be rapidly detected using molecular beacons, which fluoresce upon hybridizing to a target RNA but require oligonucleotides with complex fluorescent dye and quencher conjugations. Here, we describe a simplified method for rapid fluorescence detection of a target RNA using simple unmodified DNA oligonucleotides. To detect RNA, we developed Lettuce, a fluorogenic DNA aptamer that binds and activates the fluorescence of DFHBI-1T, an otherwise nonfluorescent molecule that resembles the chromophore found in green fluorescent protein. Lettuce was selected from a randomized DNA library based on binding to DFHBI-agarose. We further show that Lettuce can be split into two separate oligonucleotide components, which are nonfluorescent on their own but become fluorescent when their proximity is induced by a target RNA. We designed several pairs of split Lettuce fragments that contain an additional 15-20 nucleotides that are complementary to adjacent regions of the SARS-CoV-2 RNA, resulting in Lettuce fluorescence only in the presence of the viral RNA. Overall, these studies describe a simplified RNA detection approach using fully unmodified DNA oligonucleotides that reconstitute the Lettuce aptamer templated by RNA.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Aptamers, Nucleotide
/
COVID-19
Type of study:
Diagnostic study
/
Experimental Studies
/
Randomized controlled trials
Limits:
Humans
Language:
English
Journal:
ACS Chem Biol
Year:
2022
Document Type:
Article
Affiliation country:
Acschembio.1c00893
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