Naturally occurring three-way junctions can be repurposed as genetically encoded RNA-based sensors.
| Publication Type | Academic Article |
| Authors | Moon J, Wu J, Dey S, Litke J, Li X, Kim H, Jaffrey S |
| Journal | Cell Chem Biol |
| Volume | 28 |
| Issue | 11 |
| Pagination | 1569-1580.e4 |
| Date Published | 05/18/2021 |
| ISSN | 2451-9448 |
| Keywords | Aptamers, Nucleotide, Biosensing Techniques, Fluorescent Dyes, RNA, Small Molecule Libraries |
| Abstract | Small molecules can be imaged in living cells using biosensors composed of RNA. However, RNA-based devices are difficult to design. Here, we describe a versatile platform for designing RNA-based fluorescent small-molecule sensors using naturally occurring highly stable three-way junction RNAs. We show that ligand-binding aptamers and fluorogenic aptamers can be inserted into three-way junctions and connected in a way that enables the three-way junction to function as a small-molecule-regulated fluorescent sensor in vitro and in cells. The sensors are designed so that the interhelical stabilizing interactions in the three-way junction are only induced upon ligand binding. We use these RNA-based devices to measure the dynamics of S-adenosylmethionine levels in mammalian cells in real time. We show that this strategy is compatible with diverse metabolite-binding RNA aptamers, fluorogenic aptamers, and three-way junctions. Overall, these data demonstrate a versatile method for readily generating RNA devices that function in living cells. |
| DOI | 10.1016/j.chembiol.2021.04.022 |
| PubMed ID | 34010626 |
| PubMed Central ID | PMC8573057 |