biostudies-literatureUnknownBerndt ANIBIB NIH HHSNIDA NIH HHSNIA NIH HHSNIMH NIH HHSNINDS NIH HHSNIAMS NIH HHSNIGMS NIH HHSH₂O₂ is a key oxidant in mammalian biology and a pleiotropic signaling molecule at the physiological level, and its excessive accumulation in conjunction with decreased cellular reduction capacity is often found to be a common pathological marker. Here, we present a red fluorescent Genetically Encoded H₂O₂ Indicator (GEHI) allowing versatile optogenetic dissection of redox biology. Our new GEHI, oROS-HT, is a chemigenetic sensor utilizing a HaloTag and Janelia Fluor (JF) rhodamine dye as fluorescent reporters. We developed oROS-HT through a structure-guided approach aided by classic protein structures and recent protein structure prediction tools. Optimized with JF₆₃₅, oROS-HT is a sensor with 635 nm excitation and 650 nm emission peakResearch squarers.3.rs-3974015https://www.ebi.ac.uk/biostudies/studies/S-EPMC11065073biostudies-literatureFar-red and sensitive sensor for monitoring real time H<sub>2</sub>O<sub>2</sub> dynamics with subcellular resolution and in multi-parametric imaging applications.PMC11065073RM1 GM131981R01 GM139850RF1 MH130391U01 NS128537T32 AG066574R21 DA051193F31 DA056121T32 EB032787P30 DA048736P30 AR074990Moghadasi ASmith ALee JMack DNance EBremner SZuniga SWilliams AJin ZGibbs CEvitts KChavan VWait SBoyle PMoussavi-Harami FBerndt AAsencio ARegnier MNguyen AYoung JfalseFar-red and sensitive sensor for monitoring real time H<sub>2</sub>O<sub>2</sub> dynamics with subcellular resolution and in multi-parametric imaging applications.H₂O₂ is a key oxidant in mammalian biology and a pleiotropic signaling molecule at the physiological level, and its excessive accumulation in conjunction with decreased cellular reduction capacity is often found to be a common pathological marker. Here, we present a red fluorescent Genetically Encoded H₂O₂ Indicator (GEHI) allowing versatile optogenetic dissection of redox biology. Our new GEHI, oROS-HT, is a chemigenetic sensor utilizing a HaloTag and Janelia Fluor (JF) rhodamine dye as fluorescent reporters. We developed oROS-HT through a structure-guided approach aided by classic protein structures and recent protein structure prediction tools. Optimized with JF₆₃₅, oROS-HT is a sensor with 635 nm excitation and 650 nm emission peak2024-01-01T00:00:00Z2024 Apr2026-04-13T03:18:45.013Z2026-04-13T03:12:20.676ZS-EPMC110650733869933210.21203/rs.3.rs-3974015/v1