@phdthesis{oai:oist.repo.nii.ac.jp:00002804,
 author = {Tabuchi Yagui, Takeshi Ricardo},
 month = {2022-10-03, 2022-10-03},
 note = {Riboswitches have recently attracted the attention of synthetic biologists as an alternative to transcription factors for genetic regulation because of their engineerability, relative simplicity, and potential for responding to a wide array of chemical signals. However, biological constraints such as cell permeability, metabolic stability, and toxicity of their chemical ligands have prevented the development of some of those devices using conventional approaches with living cells. Cell-free systems are generally not subject to such constraints and offer a unique platform for building biochemical and genetic systems that display complex functions without using living cells. Efforts to engineer regulatory components directly in cell-free systems thus far have been based on low-throughput experimental approaches, limiting the availability of basic components for building genetically programmed cell-free systems. Here, I report a high-throughput screening method for engineering riboswitches directly in a cell-free system. Fluorescence-activated droplet-sorting (FADS) of randomized riboswitch libraries in a cell-free protein synthesis (CFPS) system rapidly identified cell-free riboswitches that respond to histamine and ciprofloxacin, compounds that are normally not compatible with conventional bacterial screening methods. Finally, the riboswitches obtained through this method were used to demonstrate chemical communication between microdroplets in a self-contained system.},
 school = {Okinawa Institute of Science and Technology Graduate University},
 title = {高スループットスクリーニングによるセルフリーリボスイッチの開発と微小液滴間化学通信への応用},
 year = {}
}