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Nat Methods ; 14(4): 391-394, 2017 Apr.
Article in English | MEDLINE | ID: mdl-28288123

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.


Subject(s)
Optogenetics/methods , Protein Engineering/methods , Recombinant Proteins/metabolism , Connexins/genetics , Connexins/metabolism , Directed Molecular Evolution , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , HEK293 Cells , HSP90 Heat-Shock Proteins/genetics , HSP90 Heat-Shock Proteins/metabolism , Humans , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Nuclear Localization Signals/genetics , Patch-Clamp Techniques , Photochemistry/methods , Recombinant Proteins/genetics , Viral Nonstructural Proteins/genetics , Viral Nonstructural Proteins/metabolism , Red Fluorescent Protein
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