PT  - JOURNAL ARTICLE
AU  - Torelli, Emanuela
AU  - Shirt-Ediss, Benjamin
AU  - Navarro, Silvia A.
AU  - Manzano, Marisa
AU  - Vizzini, Priya
AU  - Krasnogor, Natalio
TI  - Light-up split Broccoli aptamer as a versatile tool for RNA assembly monitoring in cell-free TX-TL system, hybrid RNA/DNA origami tagging and DNA biosensing
AID  - 10.1101/2022.07.20.500791
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.07.20.500791
4099  - http://biorxiv.org/content/early/2022/07/21/2022.07.20.500791.short
4100  - http://biorxiv.org/content/early/2022/07/21/2022.07.20.500791.full
AB  - Binary light-up aptamers are intriguing and emerging tools with potential in different fields. Herein, we demonstrate the versatility of a split Broccoli aptamer system able to turn on the fluorescence signal only in the presence of a complementary sequence.First, an RNA three-way junction harbouring the split system was assembled in an E. coli based cell-free TX-TL system where the folding of the functional aptamer is demonstrated. Then, the same strategy is introduced into a ‘bio-orthogonal’ hybrid RNA/DNA rectangle origami characterized by atomic force microscopy: the activation of the split system through the origami self-assembly is demonstrated. Finally, our system is successfully used to detect femtomoles of a Campylobacter spp. DNA target sequence.Potential applications of our system include real-time monitoring of the self-assembly of nucleic acid-based devices in vivo and of intracellular delivery of therapeutic nanostructures, as well as in vitro and in vivo detection of different DNA/RNA target.Competing Interest StatementThe authors have declared no competing interest.