Bioluminescent optogenetics (BL-OG) uses light emitted from a luciferase in the presence of its luciferin to activate light sensing molecules, including channelrhodopsins and light-sensitive transcription factors. Modifications to all components utilized in this approach, luciferases, luciferins, and light sensors, can aid in further research by improving, refining, and expanding the optogenetic toolbox. While luciferins are being modified chemically, light emitters and sensors can be improved by molecular evolution. Depending on available information about the protein structure, an initial step is to introduce mutations in a directed fashion. Alternatively, molecular evolution can proceed immediately or following rational mutagenesis, through random mutagenesis. We have developed a workflow for molecular evolution in mammalian cells, as the BL-OG approach is designed to be applied in mammalian brains. We tested the initial steps of molecular evolution using the fluorescent protein GFP.
Notes:
This research was supported by the the National Science Foundation (NSF) and its Division of Biological Infrastructure under Award No. 1707352
Simkins, Jacob, Crespo, Emmanuel L., and Hochgeschwender, Ute,
"Molecular Evolution of BL-OG Components"
(2022).
NeuroNex Data and Research Products.
Brown Digital Repository. Brown University Library.
https://doi.org/10.26300/adwp-fg70
This collection contains publicly available research products, including data, codes, and publications, among others. The NeuroNex Technology Hub is supported by the National Science Foundation (NSF) and its Division of Biological Infrastructure under Award No. 1707352.
