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Yet, the chromophore responsible for the fluorescence of the GFP is not emissive when isolated in solution, outside the protein environment. The most accepted explanation is that the quenching of the fluorescence results from the rotation of the aryl\u2013alkene bond and from the Z\/E isomerization. Over the years, many efforts have been performed to block these torsional rotations, mimicking the environment inside the protein \u03b2-barrel, to restore the emission intensity. Molecule rigidification through chemical modifications or complexation, or through crystallization, is one of the strategies used. This review presents an overview of the strategies developed to achieve highly emissive GFP chromophore by hindering the torsional rotations.<\/jats:p>","DOI":"10.3390\/molecules28010234","type":"journal-article","created":{"date-parts":[[2022,12,28]],"date-time":"2022-12-28T05:31:54Z","timestamp":1672205514000},"page":"234","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Locking the GFP Fluorophore to Enhance Its Emission Intensity"],"prefix":"10.3390","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0995-8680","authenticated-orcid":false,"given":"Joana R. M.","family":"Ferreira","sequence":"first","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry, University of Aveiro, Campus de Santiago, 3010-193 Aveiro, Portugal"}]},{"given":"C\u00e1tia I. C.","family":"Esteves","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry, University of Aveiro, Campus de Santiago, 3010-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6712-752X","authenticated-orcid":false,"given":"Maria Manuel B.","family":"Marques","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry, School of Science and Technology, New University of Lisbon, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7439-209X","authenticated-orcid":false,"given":"Samuel","family":"Guieu","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry, University of Aveiro, Campus de Santiago, 3010-193 Aveiro, Portugal"},{"name":"CICECO-Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, Campus de Santiago, 3010-193 Aveiro, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Omary, M.A., and Patterson, H.H. 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