Experiment Overview
| Repository ID: | FR-FCM-Z3KY | Experiment name: | De novo generated toehold switch endogenous RNA sensors in vivo assays_Expt#4 | MIFlowCyt score: | 41.00% |
| Primary researcher: | Gerard Minuesa | PI/manager: | Gerard Minuesa | Uploaded by: | Gerard Minuesa |
| Experiment dates: | 2020-10-06 - | Dataset uploaded: | Mar 2021 | Last updated: | Mar 2021 |
| Keywords: | [riboswitches] [RNA sensors] | Manuscripts: | [PMC8136780] | ||
| Organizations: | None | ||||
| Purpose: | We aimed to test endogenous RNA regulation via a toehold switch sensor to detect the E.coli small RNA RyhB, a small transcript that down-regulates a series of iron-associated genes when iron endogenous levels are low. To do so, E.coli cells constitutively expressing RyhB-responsive toehold switches (RyhB control and RyhB Moirai BD1-3) cultured in LB and 100 microM FeSO4 were induced by chelating iron with 2,2-bipyridyl at different concentrations (0 to 0.6 mM) during 1 h and the GFPmut3b-ASV expression was followed by flow cytometry (FL-1 channel). | ||||
| Conclusion: | RyhB Moirai RNA switches can optimally sense ryhB sRNA in the presence of 2,2-bipyridil increasing concentrations up to 0.3 mM (plateau). | ||||
| Comments: | Experiment #4 out of 4 in total. Legend from samples: 'bp' = 2,2-bibyridil Experiments performed by Dr. Cristina Alsina and Dr. Gerard Minuesa under supervision of Dr. Ivan Dotu at Moirai Biodesign in Barcelona Scientific Park. | ||||
| Funding: | The research conducted by Gerard Minuesa leading to these results has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under the Marie Sklodowska-Curie Grant agreement no. TECSPR-18-1-0035 (TECNIOSpring Plus) and from the Agency for Business and Competitiveness (ACCIÓ) of the Government of Catalonia. | ||||
| Quality control: | None | ||||
