Biology subjects

Verkhusha, V. V.

Publications and source records attributed to Verkhusha, V. V..

2 recordsLinked to original sources

bioRxiv2026

Compact red-shifted near-infrared fluorescent proteins enable deep-tissue SWIR imaging with in vivo optical clearing.

Compact fluorescent proteins (FPs) with red-shifted emission are needed for deep-tissue short-wavelength infrared (SWIR) imaging. We engineered a GAF domain from the JSC1 cyanobacteriochrome of thermophilic Leptolyngbya sp. into three monomeric, biliverdin-binding FPs of 19.1 kDa: miRFP729nano, miRFP732nano and miRFP735nano, with excitation/emission maxima of 714/729, 716/732 and 719/735 nm, respectively. Their off-peak fluorescence beyond 1,000 nm was several-fold higher than that of miRFP718nano previously used for SWIR imaging. miRFP732nano functioned as a fusion tag, a component of target-stabilized nanobodies, and a reporter of NF-{kappa}B and AP-1 transcriptional activities. It enabled single-laser, dual-color three-photon imaging with EGFP to depths of [~]950 m in cortex and [~]300 m in spinal cord. In mice, miRFP732nano supported SWIR imaging of skeletal muscle, inflammatory signaling and intracellular targets. Combining SWIR detection with biocompatible 4-aminoantipyrine-based in vivo tissue clearing enhanced signal and image sharpness. These red-shifted NIR FPs expand the genetically encoded toolkit for deep-tissue imaging.

bioengineering↗

bioRxiv2025

Synthetic multicolor antigen-stabilizable nanobody platform for intersectional labelling and functional imaging.

We present a synthetic toolkit of antigen-stabilizable fluorescent nanobodies (VIS-Fbs) spanning the entire visible spectrum from 450 nm to 660 nm. By engineering over twenty fluorescent proteins (FPs) and biosensors into eight nanobodies, we established a generalizable design of VIS-Fbs, which fluoresce brightly only upon binding to cognate antigens. Our synthetic approach includes constitutive, photoactivatable and photoswitchable FPs, and intensiometric FP-based biosensors. VIS-Fbs carrying biosensors enable simultaneous monitoring of two metabolites at confined locations, while FP-based VIS-Fbs targeting biosensors allow ratiometric functional imaging in the mouse brain. We further used VIS-Fbs to track endogenous {beta}-catenin dynamics in zebrafish embryos during normal development and under Wnt/{beta}-catenin signaling modulation. VIS-Fbs provide background-free visualization of intracellular proteins, multicolor detection of multiple antigens, and selective targeting of defined cell populations and compartments. This synthetic biology-driven platform enables precise studies of protein dynamics, cellular processes, and complex biological systems with high specificity and minimal background.

synthetic biology↗