Repurposing an adenine riboswitch into a fluorogenic imaging and sensing tag Journal Article
| Authors: | Dey, S. K.; Filonov, G. S.; Olarerin-George, A. O.; Jackson, B. T.; Finley, L. W. S.; Jaffrey, S. R. |
| Article Title: | Repurposing an adenine riboswitch into a fluorogenic imaging and sensing tag |
| Abstract: | Fluorogenic RNA aptamers are used to genetically encode fluorescent RNA and to construct RNA-based metabolite sensors. Unlike naturally occurring aptamers that efficiently fold and undergo metabolite-induced conformational changes, fluorogenic aptamers can exhibit poor folding, which limits their cellular fluorescence. To overcome this, we evolved a naturally occurring well-folded adenine riboswitch into a fluorogenic aptamer. We generated a library of roughly 1015 adenine aptamer-like RNAs in which the adenine-binding pocket was randomized for both size and sequence, and selected Squash, which binds and activates the fluorescence of green fluorescent protein-like fluorophores. Squash exhibits markedly improved in-cell folding and highly efficient metabolite-dependent folding when fused to a S-adenosylmethionine (SAM)-binding aptamer. A Squash-based ratiometric sensor achieved quantitative SAM measurements, revealed cell-to-cell heterogeneity in SAM levels and revealed metabolic origins of SAM. These studies show that the efficient folding of naturally occurring aptamers can be exploited to engineer well-folded cell-compatible fluorogenic aptamers and devices. [Figure not available: see fulltext.] © 2021, The Author(s), under exclusive licence to Springer Nature America, Inc. |
| Keywords: | mouse; animal; animals; mice; cells, cultured; embryonic stem cell; fluorescent dye; fluorescent dyes; chemistry; cell culture; embryonic stem cells; embryo, mammalian; conformation; nucleic acid conformation; genetic procedures; fluorescence imaging; aptamer; aptamers, nucleotide; optical imaging; procedures; biosensing techniques; mammalian embryo; humans; human; male |
| Journal Title: | Nature Chemical Biology |
| Volume: | 18 |
| Issue: | 2 |
| ISSN: | 1552-4450 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2022-02-01 |
| Start Page: | 180 |
| End Page: | 190 |
| Language: | English |
| DOI: | 10.1038/s41589-021-00925-0 |
| PUBMED: | 34937909 |
| PROVIDER: | scopus |
| PMCID: | PMC8967656 |
| DOI/URL: | |
| Notes: | Article -- Erratum issued, see DOI: 10.1038/s41589-022-00969-w -- Export Date: 1 March 2022 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work