Small near-infrared photochromic protein for photoacoustic multi-contrast imaging and detection of protein interactions in vivo

Nature Communications
Lei LiVladislav V Verkhusha

Abstract

Photoacoustic (PA) computed tomography (PACT) benefits from genetically encoded probes with photochromic behavior, which dramatically increase detection sensitivity and specificity through photoswitching and differential imaging. Starting with a DrBphP bacterial phytochrome, we have engineered a near-infrared photochromic probe, DrBphP-PCM, which is superior to the full-length RpBphP1 phytochrome previously used in differential PACT. DrBphP-PCM has a smaller size, better folding, and higher photoswitching contrast. We have imaged both DrBphP-PCM and RpBphP1 simultaneously on the basis of their unique signal decay characteristics, using a reversibly switchable single-impulse panoramic PACT (RS-SIP-PACT) with a single wavelength excitation. The simple structural organization of DrBphP-PCM allows engineering a bimolecular PA complementation reporter, a split version of DrBphP-PCM, termed DrSplit. DrSplit enables PA detection of protein-protein interactions in deep-seated mouse tumors and livers, achieving 125-µm spatial resolution and 530-cell sensitivity in vivo. The combination of RS-SIP-PACT with DrBphP-PCM and DrSplit holds great potential for noninvasive multi-contrast deep-tissue functional imaging.

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Citations

Dec 18, 2018·Advanced Materials·Qinrui FuXiaoyuan Chen
Jun 20, 2019·Biochemistry. Biokhimii︠a︡·M M KarasevV V Verkhusha
Jun 26, 2020·Science Advances·Kanuj MishraAndre C Stiel
Feb 18, 2021·European Journal of Nuclear Medicine and Molecular Imaging·Daniel RazanskyRuiqing Ni
Feb 25, 2021·Scientific Reports·Matthew D LaramieMaged Henary
Jan 26, 2021·Medical Physics·Hongjin XiangHuabei Jiang
Apr 10, 2021·The Journal of Chemical Physics·Dongkwan LeeLu Wei
Jul 26, 2018·Trends in Biotechnology·Daria M ShcherbakovaVladislav V Verkhusha
May 21, 2021·Nanoscale Research Letters·Zhongyang YuKaiwen Hu
Aug 8, 2019·ACS Chemical Biology·Juan Pablo Fuenzalida WernerAndre C Stiel
Apr 2, 2019·Analytical Chemistry·Kanuj MishraAndre C Stiel

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Methods Mentioned

BETA
FRET
BRET
protein folding
transfection
PCR
fluorescence
flow
flow cytometry
fluorescence imaging

Software Mentioned

Rapa
PACT
BiFC
PCM
DrSplit
RS
DrBphP
SIP
Sim
ARNT

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