Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Optogenetics enables the spatio-temporally precise control of cell and animal behavior. Many optogenetic tools are driven by light-controlled protein–protein interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs). Applying light-controlled PPIs to new target proteins is challenging because it is difficult to predict which of the many available LSDs, if any, will yield robust light regulation. As a consequence, fusion protein libraries need to be prepared and tested, but methods and platforms to facilitate this process are currently not available. Here, we developed a genetic engineering strategy and vector library for the rapid generation of light-controlled PPIs. The strategy permits fusing a target protein to multiple LSDs efficiently and in two orientations. The public and expandable library contains 29 vectors with blue, green or red light-responsive LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate the versatility of the approach and the necessity for sampling LSDs by generating light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a new resource for optical regulation of a broad range of target proteins in cell and developmental biology.

Original languageEnglish
Pages (from-to)3046-3055
Number of pages10
JournalJournal of Molecular Biology
Volume431
Issue number17
DOIs
Publication statusPublished - 9 Aug 2019

Keywords

  • Optogenetics
  • light-sensitive domain
  • photoreceptor
  • protein-protein interaction
  • caspase9

Cite this

@article{f6d12fdbc83c47019f3614fda70c3575,
title = "Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions",
abstract = "Optogenetics enables the spatio-temporally precise control of cell and animal behavior. Many optogenetic tools are driven by light-controlled protein–protein interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs). Applying light-controlled PPIs to new target proteins is challenging because it is difficult to predict which of the many available LSDs, if any, will yield robust light regulation. As a consequence, fusion protein libraries need to be prepared and tested, but methods and platforms to facilitate this process are currently not available. Here, we developed a genetic engineering strategy and vector library for the rapid generation of light-controlled PPIs. The strategy permits fusing a target protein to multiple LSDs efficiently and in two orientations. The public and expandable library contains 29 vectors with blue, green or red light-responsive LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate the versatility of the approach and the necessity for sampling LSDs by generating light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a new resource for optical regulation of a broad range of target proteins in cell and developmental biology.",
keywords = "Optogenetics, light-sensitive domain, photoreceptor, protein-protein interaction, caspase9",
author = "Alexandra-Madelaine Tichy and Gerrard, {Elliot J.} and Legrand, {Julien M.D.} and Hobbs, {Robin M.} and Harald Janovjak",
year = "2019",
month = "8",
day = "9",
doi = "10.1016/j.jmb.2019.05.033",
language = "English",
volume = "431",
pages = "3046--3055",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Elsevier",
number = "17",

}

Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions. / Tichy, Alexandra-Madelaine; Gerrard, Elliot J.; Legrand, Julien M.D.; Hobbs, Robin M.; Janovjak, Harald.

In: Journal of Molecular Biology, Vol. 431, No. 17, 09.08.2019, p. 3046-3055.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions

AU - Tichy, Alexandra-Madelaine

AU - Gerrard, Elliot J.

AU - Legrand, Julien M.D.

AU - Hobbs, Robin M.

AU - Janovjak, Harald

PY - 2019/8/9

Y1 - 2019/8/9

N2 - Optogenetics enables the spatio-temporally precise control of cell and animal behavior. Many optogenetic tools are driven by light-controlled protein–protein interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs). Applying light-controlled PPIs to new target proteins is challenging because it is difficult to predict which of the many available LSDs, if any, will yield robust light regulation. As a consequence, fusion protein libraries need to be prepared and tested, but methods and platforms to facilitate this process are currently not available. Here, we developed a genetic engineering strategy and vector library for the rapid generation of light-controlled PPIs. The strategy permits fusing a target protein to multiple LSDs efficiently and in two orientations. The public and expandable library contains 29 vectors with blue, green or red light-responsive LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate the versatility of the approach and the necessity for sampling LSDs by generating light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a new resource for optical regulation of a broad range of target proteins in cell and developmental biology.

AB - Optogenetics enables the spatio-temporally precise control of cell and animal behavior. Many optogenetic tools are driven by light-controlled protein–protein interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs). Applying light-controlled PPIs to new target proteins is challenging because it is difficult to predict which of the many available LSDs, if any, will yield robust light regulation. As a consequence, fusion protein libraries need to be prepared and tested, but methods and platforms to facilitate this process are currently not available. Here, we developed a genetic engineering strategy and vector library for the rapid generation of light-controlled PPIs. The strategy permits fusing a target protein to multiple LSDs efficiently and in two orientations. The public and expandable library contains 29 vectors with blue, green or red light-responsive LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate the versatility of the approach and the necessity for sampling LSDs by generating light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a new resource for optical regulation of a broad range of target proteins in cell and developmental biology.

KW - Optogenetics

KW - light-sensitive domain

KW - photoreceptor

KW - protein-protein interaction

KW - caspase9

UR - http://www.scopus.com/inward/record.url?scp=85066848305&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2019.05.033

DO - 10.1016/j.jmb.2019.05.033

M3 - Article

VL - 431

SP - 3046

EP - 3055

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 17

ER -