[{"article_type":"original","year":"2019","oa_version":"Preprint","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","date_updated":"2023-08-28T09:39:22Z","external_id":{"isi":["000482872100002"]},"scopus_import":"1","publication_identifier":{"issn":["00222836"],"eissn":["10898638"]},"_id":"6564","date_published":"2019-08-09T00:00:00Z","abstract":[{"text":"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.","lang":"eng"}],"issue":"17","article_processing_charge":"No","volume":431,"publication_status":"published","main_file_link":[{"url":"http://www.biorxiv.org/content/10.1101/583369v1","open_access":"1"}],"oa":1,"day":"09","author":[{"last_name":"Tichy","full_name":"Tichy, Alexandra-Madelaine","first_name":"Alexandra-Madelaine","id":"29D8BB2C-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Gerrard, Elliot J.","first_name":"Elliot J.","last_name":"Gerrard"},{"last_name":"Legrand","full_name":"Legrand, Julien M.D.","first_name":"Julien M.D."},{"last_name":"Hobbs","first_name":"Robin M.","full_name":"Hobbs, Robin M."},{"last_name":"Janovjak","full_name":"Janovjak, Harald L","first_name":"Harald L","id":"33BA6C30-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8023-9315"}],"type":"journal_article","citation":{"short":"A.-M. Tichy, E.J. Gerrard, J.M.D. Legrand, R.M. Hobbs, H.L. Janovjak, Journal of Molecular Biology 431 (2019) 3046–3055.","apa":"Tichy, A.-M., Gerrard, E. J., Legrand, J. M. D., Hobbs, R. M., &#38; Janovjak, H. L. (2019). Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions. <i>Journal of Molecular Biology</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.jmb.2019.05.033\">https://doi.org/10.1016/j.jmb.2019.05.033</a>","ama":"Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak HL. Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions. <i>Journal of Molecular Biology</i>. 2019;431(17):3046-3055. doi:<a href=\"https://doi.org/10.1016/j.jmb.2019.05.033\">10.1016/j.jmb.2019.05.033</a>","ieee":"A.-M. Tichy, E. J. Gerrard, J. M. D. Legrand, R. M. Hobbs, and H. L. Janovjak, “Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions,” <i>Journal of Molecular Biology</i>, vol. 431, no. 17. Elsevier, pp. 3046–3055, 2019.","ista":"Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak HL. 2019. Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions. Journal of Molecular Biology. 431(17), 3046–3055.","chicago":"Tichy, Alexandra-Madelaine, Elliot J. Gerrard, Julien M.D. Legrand, Robin M. Hobbs, and Harald L Janovjak. “Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions.” <i>Journal of Molecular Biology</i>. Elsevier, 2019. <a href=\"https://doi.org/10.1016/j.jmb.2019.05.033\">https://doi.org/10.1016/j.jmb.2019.05.033</a>.","mla":"Tichy, Alexandra-Madelaine, et al. “Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions.” <i>Journal of Molecular Biology</i>, vol. 431, no. 17, Elsevier, 2019, pp. 3046–55, doi:<a href=\"https://doi.org/10.1016/j.jmb.2019.05.033\">10.1016/j.jmb.2019.05.033</a>."},"title":"Engineering strategy and vector library for the rapid generation of modular light-controlled protein–protein interactions","language":[{"iso":"eng"}],"doi":"10.1016/j.jmb.2019.05.033","month":"08","date_created":"2019-06-16T21:59:14Z","page":"3046-3055","quality_controlled":"1","department":[{"_id":"HaJa"}],"publication":"Journal of Molecular Biology","status":"public","intvolume":"       431","publisher":"Elsevier","isi":1}]