Abstract
To expand the range of experiments that are accessible with optogenetics, we developed a photocleavable protein (PhoCl) that spontaneously dissociates into two fragments after violet-light-induced cleavage of a specific bond in the protein backbone. We demonstrated that PhoCl can be used to engineer light-activatable Cre recombinase, Gal4 transcription factor, and a viral protease that in turn was used to activate opening of the large-pore ion channel Pannexin-1.
MeSH terms
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Connexins / genetics
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Connexins / metabolism
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Directed Molecular Evolution
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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HEK293 Cells
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HSP90 Heat-Shock Proteins / genetics
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HSP90 Heat-Shock Proteins / metabolism
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Humans
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism
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Nuclear Localization Signals / genetics
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Optogenetics / methods*
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Patch-Clamp Techniques
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Photochemistry / methods
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Protein Engineering / methods*
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism*
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Red Fluorescent Protein
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Viral Nonstructural Proteins / genetics
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Viral Nonstructural Proteins / metabolism
Substances
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Connexins
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HSP90 Heat-Shock Proteins
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Luminescent Proteins
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NS3 protein, hepatitis C virus
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Nerve Tissue Proteins
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Nuclear Localization Signals
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PANX1 protein, human
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Recombinant Proteins
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Viral Nonstructural Proteins
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Green Fluorescent Proteins