Detection of SARS-CoV-2 RNA Using a DNA Aptamer Mimic of Green Fluorescent Protein.
| Publication Type | Academic Article |
| Authors | VarnBuhler B, Moon J, Dey S, Wu J, Jaffrey S |
| Journal | ACS Chem Biol |
| Volume | 17 |
| Issue | 4 |
| Pagination | 840-853 |
| Date Published | 03/26/2022 |
| ISSN | 1554-8937 |
| Keywords | Aptamers, Nucleotide, COVID-19 |
| 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. |
| DOI | 10.1021/acschembio.1c00893 |
| PubMed ID | 35341244 |
| PubMed Central ID | PMC9780036 |