Leong, Lee Min Kang, Bok Eum Baker, Bradley J. 2024-01-19T14:33:37Z 2024-01-19T14:33:37Z 2021-10-21 2021-05-18 0006-3495 https://pubs.kist.re.kr/handle/201004/116986 A new family of genetically encoded voltage indicators (GEVIs) has been developed based on intermolecular Forster resonance energy transfer (FRET). To test the hypothesis that the GEVI ArcLight functions via interactions between the fluorescent protein (FP) domains of neighboring probes, the FP of ArcLight was replaced with either a FRET donor or acceptor FP. We discovered relatively large FRET signals only when cells were cotransfected with both the FRET donor and acceptor GEVIs. Using a cyan fluorescent protein donor and an RFP acceptor, we were able to observe a voltage-dependent signal with an emission peak separated by over 200 nm from the excitation wavelength. The intermolecular FRET strategy also works for rhodopsin-based probes, potentially improving their flexibility as well. Separating the FRET pair into two distinct proteins has important advantages over intramolecular FRET constructs. The signals are larger because the voltage-induced conformational change moves two FPs independently. The expression of the FRET donor and acceptor can also be restricted independently, enabling greater cell type specificity as well as refined subcellular voltage reporting. English CELL PRESS FLUORESCENT PROTEIN BRIGHT NEURONS VARIANT MICE Improving the flexibility of genetically encoded voltage indicators via intermolecular FRET Article 10.1016/j.bpj.2021.03.010 1 BIOPHYSICAL JOURNAL, v.120, no.10, pp.1927 - 1941 BIOPHYSICAL JOURNAL 120 10 1927 1941 scie scopus 000652829900008 2-s2.0-85103310094 Biophysics Biophysics Article FLUORESCENT PROTEIN BRIGHT NEURONS VARIANT MICE