Design of Orthogonal Far-Red, Orange and Green Fluorophore-binding Proteins for Multiplex Imaging.

ABSTRACT: Fluorescent proteins and small molecule dyes have complementary strengths for biological imaging: the former are genetically manipulatable enabling tagging of specific proteins and detection of protein interactions, while the latter have greater photostability and brightness but are difficult to target. To combine these strengths, we used de novo protein design to generate binders to three bright, stable, cell-permeable dyes spanning the visible spectrum: JF657 (far red), JF596 (orange-red) and JF494 (green). For each dye, we obtain nanomolar binders with weak or no binding to the other two dyes; the accuracy of the design approach is confirmed by a crystal structure of one binder which is very close to the design model. Fusion of the JF567, JF596 and JF494 binders to three different targe

SUBMITTER: Tran L 

PROVIDER: S-EPMC12424662 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature