Opto-MASS: a high-throughput engineering platform for genetically encoded fluorescentsensors enabling all optical in vivo detection of monoamines and neuropeptides
Rappleye, M.; Gordon-Fennel, A.; Castro, D. C.; Matarasso, A. K.; Zamorano, C. A.; Wait, S. J.; Lee, J. D.; Siebart, J. D.; Suko, A.; Smith, N. C.; Muster, J.; Matreyek, K. A.; Fowler, D. M.; Stuber, G. D.; Bruchas, M. R.; Berndt, A.
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Fluorescent sensor proteins are instrumental for detecting biological signals in vivo with high temporal accuracy and cell-type specificity. However, engineering sensors with physiological ligand sensitivity and selectivity is difficult because they need to be optimized through individual mutagenesis in vitro to assess their performance. The vast mutational landscape proteins constitute an obstacle that slows down sensor development. This is particularly true for sensors that require mammalian host systems to be screened. Here, we developed a novel high-throughput engineering platform that functionally tests thousands of variants simultaneously in mammalian cells and thus allows the screening of large variant numbers. We showcase the capabilities of our platform, called Optogenetic Microwell Array Screening System (Opto-MASS), by engineering novel monoamine and neuropeptide in vivo capable sensors with distinct physiological roles at high-throughput.
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