Near-infrared optogenetic pair for protein regulation and spectral multiplexing

Taras A. Redchuk, Evgeniya S. Omelina, Konstantin G. Chernov, Vladislav Verkhusha

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1–PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization. We exploited a helix–PAS fold of Q-PAS1 to develop several near-infrared-light-controllable transcription regulation systems, enabling either 40-fold activation or inhibition. The light-induced BphP1–Q-PAS1 interaction allowed modification of the chromatin epigenetic state. Multiplexing the BphP1–Q-PAS1 pair with a blue-light-activatable LOV-domain-based system demonstrated their negligible spectral crosstalk. By integrating the Q-PAS1 and LOV domains in a single optogenetic tool, we achieved tridirectional protein targeting, independently controlled by near-infrared and blue light, thus demonstrating the superiority of Q-PAS1 for spectral multiplexing and engineering of multicomponent systems.

Original languageEnglish (US)
JournalNature Chemical Biology
DOIs
StateAccepted/In press - Mar 27 2017

Fingerprint

Optogenetics
Light
Proteins
Phytochrome
Protein Transport
Epigenomics
Chromatin
Biomedical Research

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Near-infrared optogenetic pair for protein regulation and spectral multiplexing. / Redchuk, Taras A.; Omelina, Evgeniya S.; Chernov, Konstantin G.; Verkhusha, Vladislav.

In: Nature Chemical Biology, 27.03.2017.

Research output: Contribution to journalArticle

@article{20f7a7c48bef4716b7c323def8b7d8c5,
title = "Near-infrared optogenetic pair for protein regulation and spectral multiplexing",
abstract = "Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1–PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization. We exploited a helix–PAS fold of Q-PAS1 to develop several near-infrared-light-controllable transcription regulation systems, enabling either 40-fold activation or inhibition. The light-induced BphP1–Q-PAS1 interaction allowed modification of the chromatin epigenetic state. Multiplexing the BphP1–Q-PAS1 pair with a blue-light-activatable LOV-domain-based system demonstrated their negligible spectral crosstalk. By integrating the Q-PAS1 and LOV domains in a single optogenetic tool, we achieved tridirectional protein targeting, independently controlled by near-infrared and blue light, thus demonstrating the superiority of Q-PAS1 for spectral multiplexing and engineering of multicomponent systems.",
author = "Redchuk, {Taras A.} and Omelina, {Evgeniya S.} and Chernov, {Konstantin G.} and Vladislav Verkhusha",
year = "2017",
month = "3",
day = "27",
doi = "10.1038/nchembio.2343",
language = "English (US)",
journal = "Nature Chemical Biology",
issn = "1552-4450",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - Near-infrared optogenetic pair for protein regulation and spectral multiplexing

AU - Redchuk, Taras A.

AU - Omelina, Evgeniya S.

AU - Chernov, Konstantin G.

AU - Verkhusha, Vladislav

PY - 2017/3/27

Y1 - 2017/3/27

N2 - Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1–PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization. We exploited a helix–PAS fold of Q-PAS1 to develop several near-infrared-light-controllable transcription regulation systems, enabling either 40-fold activation or inhibition. The light-induced BphP1–Q-PAS1 interaction allowed modification of the chromatin epigenetic state. Multiplexing the BphP1–Q-PAS1 pair with a blue-light-activatable LOV-domain-based system demonstrated their negligible spectral crosstalk. By integrating the Q-PAS1 and LOV domains in a single optogenetic tool, we achieved tridirectional protein targeting, independently controlled by near-infrared and blue light, thus demonstrating the superiority of Q-PAS1 for spectral multiplexing and engineering of multicomponent systems.

AB - Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1–PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization. We exploited a helix–PAS fold of Q-PAS1 to develop several near-infrared-light-controllable transcription regulation systems, enabling either 40-fold activation or inhibition. The light-induced BphP1–Q-PAS1 interaction allowed modification of the chromatin epigenetic state. Multiplexing the BphP1–Q-PAS1 pair with a blue-light-activatable LOV-domain-based system demonstrated their negligible spectral crosstalk. By integrating the Q-PAS1 and LOV domains in a single optogenetic tool, we achieved tridirectional protein targeting, independently controlled by near-infrared and blue light, thus demonstrating the superiority of Q-PAS1 for spectral multiplexing and engineering of multicomponent systems.

UR - http://www.scopus.com/inward/record.url?scp=85016105871&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016105871&partnerID=8YFLogxK

U2 - 10.1038/nchembio.2343

DO - 10.1038/nchembio.2343

M3 - Article

JO - Nature Chemical Biology

JF - Nature Chemical Biology

SN - 1552-4450

ER -