Design, construction, and validation of optogenetic proteins.
Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, United States.
Department of Chemistry, University of North Carolina, Chapel Hill, NC, United States; Department of Pharmacology, University of North Carolina, Chapel Hill, NC, United States. Electronic address: [email protected]
- Published Article
Methods in enzymology
- Publication Date
Jan 01, 2019
Cellular optogenetics employs light-regulated, genetically encoded protein actuators to perturb cellular signaling with unprecedented spatial and temporal control. Here, we present a potentially generalized approach for transforming a given protein of interest (POI) into an optogenetic species. We describe the rational and methods by which we developed three different optogenetic POIs utilizing the Cry2-Cib photodimerizing pair. The process pipeline is highlighted by (1) developing a low level, constitutively active POI that is independent of endogenous regulation, (2) fusion of the mutant protein of interest to an optogenetic photodimerizing system, and (3) light-mediated recruitment of the light-responsive POI to specific subcellular regions. © 2019 Elsevier Inc. All rights reserved.
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This record was last updated on 10/23/2019 and may not reflect the most current and accurate biomedical/scientific data available from NLM.
The corresponding record at NLM can be accessed at https://www.ncbi.nlm.nih.gov/pubmed/31128778