Multiplex Neural Circuit Tracing With G-Deleted Rabies Viral Vectors DOI Creative Commons
Toshiaki Suzuki,

Nao Morimoto,

Akinori Akaike

и другие.

Frontiers in Neural Circuits, Год журнала: 2020, Номер 13

Опубликована: Янв. 10, 2020

Neural circuits interconnect to organize large-scale networks that generate perception, cognition, memory, and behavior. Information in the nervous system is processed both through parallel, independent intermixing circuits. Analyzing interaction between particularly indispensable for elucidating how brain functions. Monosynaptic circuit tracing with glycoprotein (G) gene-deleted rabies viral vectors (RVdG) comprises a powerful approach studying structure function of neural Pseudotyping RVdG foreign envelope EnvA permits expression transgenes such as fluorescent proteins, genetically-encoded sensors, or optogenetic tools cells expressing TVA, cognate receptor EnvA. Trans-complementation virus glycoproteins (RV-G) enables trans-synaptic labeling input neurons directly connected starter TVA RV-G. However, it remains challenging simultaneously map neuronal connections from multiple cell populations their interactions solely EnvA/TVA-mediated RV single animal. To overcome this limitation, here, we multiplexed by optimizing distinct envelopes (oEnvX) corresponding receptors (oTVX). Based on EnvB/TVB EnvE/DR46-TVB systems derived avian sarcoma leukosis virus, developed optimized TVB lower higher affinity (oTVB-L oTVB-H) chimeric oEnvB, well an TVE (oTVE-H) its oEnvE. We demonstrated independence infection oEnvA/oTVA, oEnvB/oTVB, oEnvE/oTVE vivo proof-of-concept multiplex two classes layer 5 targeting either other cortical subcortical areas. also successfully labeled common lateral geniculate nucleus cortico-cortical inhibitory mouse V1 tracing. These allow differential uncover mechanisms underlying parallel processing integrated brain.

Язык: Английский

Temporally multiplexed imaging of dynamic signaling networks in living cells DOI Creative Commons
Yong Qian,

Orhan T. Celiker,

Zeguan Wang

и другие.

Cell, Год журнала: 2023, Номер 186(25), С. 5656 - 5672.e21

Опубликована: Ноя. 28, 2023

Molecular signals interact in networks to mediate biological processes. To analyze these networks, it would be useful image many at once, the same living cell, using standard microscopes and genetically encoded fluorescent reporters. Here, we report temporally multiplexed imaging (TMI), which uses proteins with different clocklike properties—such as reversibly photoswitchable switching kinetics—to represent cellular signals. We linearly decompose a brief (few-second-long) trace of fluorescence fluctuations, each point into weighted sum traces exhibited by fluorophore expressed cell. The weights then signal amplitudes. use TMI relationships between kinase activities individual cells, well cell-cycle signals, pointing toward broad utility throughout biology analysis transduction cascades systems.

Язык: Английский

Процитировано

25
Fluorescent Biosensors for Neuronal Metabolism and the Challenges of Quantitation DOI
Dorothy Koveal, Carlos Manlio Díaz‐García, Gary Yellen

и другие.

Current Opinion in Neurobiology, Год журнала: 2020, Номер 63, С. 111 - 121

Опубликована: Июнь 16, 2020

Язык: Английский

Процитировано

55
Single-component near-infrared optogenetic systems for gene transcription regulation DOI Creative Commons
Andrii A. Kaberniuk, Mikhail Baloban, Mikhail Monakhov

и другие.

Nature Communications, Год журнала: 2021, Номер 12(1)

Опубликована: Июнь 23, 2021

Abstract Near-infrared (NIR) optogenetic systems for transcription regulation are in high demand because NIR light exhibits low phototoxicity, scattering, and allows combining with probes of visible range. However, available consist several protein components large size multidomain structure. Here, we engineer single-component consisting evolved photosensory core module Idiomarina sp . bacterial phytochrome, named iLight, which smaller packable adeno-associated virus. We characterize iLight vitro gene repression activation mammalian cells. Bacterial system shows 115-fold production. Comparing to multi-component systems, higher 65-fold cells faster 6-fold deep tissues mice. Neurons transduced viral-encoded exhibit 50-fold induction fluorescent reporter. light-induced neuronal expression green-light-activatable CheRiff channelrhodopsin causes 20-fold increase photocurrent demonstrates efficient spectral multiplexing.

Язык: Английский

Процитировано

54
Booster, a Red-Shifted Genetically Encoded Förster Resonance Energy Transfer (FRET) Biosensor Compatible with Cyan Fluorescent Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue Light-Responsive Optogenetic Tools DOI
Tetsuya Watabe, Kenta Terai, Kenta Sumiyama

и другие.

ACS Sensors, Год журнала: 2020, Номер 5(3), С. 719 - 730

Опубликована: Фев. 26, 2020

Genetically encoded Förster resonance energy transfer (FRET)-based biosensors have been developed for the visualization of signaling molecule activities. Currently, most them are comprised cyan and yellow fluorescent proteins (CFP YFP), precluding use multiple FRET within a single cell. Moreover, based on CFP YFP incompatible with optogenetic tools that operate at blue light. To overcome these problems, here, we red-shifted excitation emission wavelengths. We chose mKOκ mKate2 as favorable donor acceptor pair by calculating distance. By optimizing order modulatory domains biosensors, biosensor backbone named "Booster". The performance protein kinase A (PKA) Booster (Booster-PKA) was comparable to AKAR3EV, previously comprising YFP. For proof concept, first showed simultaneous monitoring activities two kinases Booster-PKA ERK Second, PKA activation Beggiatoa photoactivated adenylyl cyclase, an generator cyclic AMP. Finally, presented activity in living tissues transgenic mice expressing Booster-PKA. Collectively, results demonstrate effectiveness versatility imaging tool vitro vivo.

Язык: Английский

Процитировано

52
The Red Edge: Bilin-Binding Photoreceptors as Optogenetic Tools and Fluorescence Reporters DOI Creative Commons
Kun Tang, Hannes M. Beyer, Matías D. Zurbriggen

и другие.

Chemical Reviews, Год журнала: 2021, Номер 121(24), С. 14906 - 14956

Опубликована: Окт. 20, 2021

This review adds the bilin-binding phytochromes to Chemical Reviews thematic issue "Optogenetics and Photopharmacology". The work is structured into two parts. We first outline photochemistry of covalently bound tetrapyrrole chromophore summarize relevant spectroscopic, kinetic, biochemical, physiological properties different families phytochromes. Based on this knowledge, we then describe engineering further improve these chromoproteins as photoswitches their employment in an ever-growing number optogenetic applications. Most applications rely light-controlled complex formation between plant photoreceptor PhyB phytochrome-interacting factors (PIFs) or C-terminal light-regulated domains with enzymatic functions present many bacterial algal Phytochrome-based tools are currently implemented bacteria, yeast, plants, animals achieve light control a wide range biological activities. These cover regulation gene expression, protein transport cell organelles, recruitment phytochrome- PIF-tagged proteins membranes other cellular compartments. compilation illustrates intrinsic advantages compared classes, e.g., bidirectional dual-wavelength enabling instant ON OFF regulation. In particular, long wavelength absorption fluorescence within "transparent window" makes attractive for requiring deep tissue penetration combination blue UV light-sensing photoreceptors. addition variability employing natural engineered phytochromes, also discuss recent progress development bilin-based fluorescent proteins.

Язык: Английский

Процитировано

44
Multiphoton intravital microscopy of rodents DOI
Colinda L. G. J. Scheele, David Herrmann, Erika Yamashita

и другие.

Nature Reviews Methods Primers, Год журнала: 2022, Номер 2(1)

Опубликована: Ноя. 10, 2022

Язык: Английский

Процитировано

37
Deep-tissue SWIR imaging using rationally designed small red-shifted near-infrared fluorescent protein DOI
Olena S. Oliinyk,

Chenshuo Ma,

Sergei Pletnev

и другие.

Nature Methods, Год журнала: 2022, Номер 20(1), С. 70 - 74

Опубликована: Дек. 1, 2022

Язык: Английский

Процитировано

37
Optogenetics Illuminates Applications in Microbial Engineering DOI

Shannon M. Hoffman,

Allison Y. Tang, José L. Avalos‬

и другие.

Annual Review of Chemical and Biomolecular Engineering, Год журнала: 2022, Номер 13(1), С. 373 - 403

Опубликована: Март 23, 2022

Optogenetics has been used in a variety of microbial engineering applications, such as chemical and protein production, studies cell physiology, engineered microbe-host interactions. These diverse applications benefit from the precise spatiotemporal control that light affords, well its tunability, reversibility, orthogonality. This combination unique capabilities enabled surge recent years investigating complex biological systems with completely new approaches. We briefly describe optogenetic tools have developed for engineering, emphasizing scientific advancements they enabled. In particular, we focus on benefits implementing control, bacterial therapeutics to cybergenetics. Finally, discuss future research directions, special attention given development orthogonal multichromatic controls. With an abundance advantages offered by optogenetics, is bright engineering.

Язык: Английский

Процитировано

33
Optogenetic manipulation and photoacoustic imaging using a near-infrared transgenic mouse model DOI Creative Commons
L.A. Kasatkina,

Chenshuo Ma,

Mikhail E. Matlashov

и другие.

Nature Communications, Год журнала: 2022, Номер 13(1)

Опубликована: Май 19, 2022

Optogenetic manipulation and optical imaging in the near-infrared range allow non-invasive light-control readout of cellular organismal processes deep tissues vivo. Here, we exploit advantages Rhodopseudomonas palustris BphP1 bacterial phytochrome, which incorporates biliverdin chromophore reversibly photoswitches between ground (740-800 nm) activated (620-680 states, to generate a loxP-BphP1 transgenic mouse model. The enables Cre-dependent temporal spatial targeting expression We validate optogenetic performance endogenous BphP1, state binds its engineered protein partner QPAS1, trigger gene transcription primary cells living mice. demonstrate photoacoustic tomography different organs, developing embryos, virus-infected regenerating livers, with centimeter penetration depth. model provides opportunities for both optogenetics vivo serves as source genomically encoded BphP1.

Язык: Английский

Процитировано

33
Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report DOI Creative Commons
Ahmed S. Abdelfattah,

Srinivasa Rao Allu,

Robert E. Campbell

и другие.

Neurophotonics, Год журнала: 2022, Номер 9(S1)

Опубликована: Апрель 27, 2022

Neurophotonics was launched in 2014 coinciding with the launch of BRAIN Initiative focused on development technologies for advancement neuroscience. For last seven years, Neurophotonics' agenda has been well aligned this focus neurotechnologies featuring new optical methods and tools applicable to brain studies. While 2.0 is pivoting towards applications these novel quest understand brain, article we review an extensive diverse toolkit explore function that have emerged from related large-scale efforts measurement manipulation structure function. Here, neurophotonic mostly animal A companion article, scheduled appear later year, will cover diffuse imaging noninvasive human each domain, outline current state-of-the-art respective technologies, identify areas where innovation needed provide outlook future directions.

Язык: Английский

Процитировано

33