Optogenetic Engineering: Light‐Directed Cell Motility |
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Authors: | Prof. Dr. Robert M. Hughes Prof. Dr. David S. Lawrence |
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Affiliation: | Department of Chemistry, Division of Chemical Biology and Medicinal Chemistry, and Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599 (USA) |
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Abstract: | Genetically encoded, light‐activatable proteins provide the means to probe biochemical pathways at specific subcellular locations with exquisite temporal control. However, engineering these systems in order to provide a dramatic jump in localized activity, while retaining a low dark‐state background remains a significant challenge. When placed within the framework of a genetically encodable, light‐activatable heterodimerizer system, the actin‐remodelling protein cofilin induces dramatic changes in the F‐actin network and consequent cell motility upon illumination. We demonstrate that the use of a partially impaired mutant of cofilin is critical for maintaining low background activity in the dark. We also show that light‐directed recruitment of the reduced activity cofilin mutants to the cytoskeleton is sufficient to induce F‐actin remodeling, formation of filopodia, and directed cell motility. |
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Keywords: | cell motility cofilin F‐actin optogenetics protein design |
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