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Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions

Tichy, Alexandra-Madelaine et al.

Journal of molecular biology. Volume 431:Number 17 (2019, August 9th); pp 3046-3055 -- Elsevier Science Ltd

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  • Title:
    Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions
  • Author: Tichy, Alexandra-Madelaine;
    Gerrard, Elliot J.;
    Legrand, Julien M.D.;
    Hobbs, Robin M.;
    Janovjak, Harald
  • Found In: Journal of molecular biology. Volume 431:Number 17 (2019, August 9th); pp 3046-3055
  • Journal Title: Journal of molecular biology
  • Subjects: Biochimie--Périodiques; Biologie moléculaire--Périodiques; Biologie--Périodiques; Bacteriology--Periodicals; Molecular Biology--Periodicals; Biochemistry--Periodicals; Biology--Periodicals; Molecular biology--Periodicals; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; LSD--LOV--CRY--PHY--CBD--PPI--PHR--P2A--PCR--RT--casp9; Optogenetics--light-sensitive domain--photoreceptor--protein-protein interaction--caspase9; Dewey: 572.805
  • Rights: Licensed
  • Publication Details: Elsevier Science Ltd
  • Abstract: 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. Graphical abstract Unlabelled Image Highlights Optogenetic tools permit the spatio-temporal control of protein and cell function. A new method efficiently prepares modular optogenetic tools. The method combines a genetic engineering strategy and photoreceptor library. Using the strategy and library, light-activated caspases were developed. Optical regulation of diverse target proteins is now more directly accessible.
  • Identifier: System Number: ETOCvdc_100086797799.0x000001; Journal ISSN: 0022-2836; 10.1016/j.jmb.2019.05.033
  • Publication Date: 2019
  • Physical Description: Electronic
  • Shelfmark(s): 5020.700000
  • UIN: ETOCvdc_100086797799.0x000001

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