{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:49:07Z","timestamp":1787028547187,"version":"build-2736575974"},"reference-count":48,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2024,8,31]],"date-time":"2024-08-31T00:00:00Z","timestamp":1725062400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Michael J. Fox Foundation for Parkinson\u2019s disease","award":["MJFF-020717"],"award-info":[{"award-number":["MJFF-020717"]}]},{"name":"Government of Canada","award":["MJFF-020717"],"award-info":[{"award-number":["MJFF-020717"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>With emerging genetic association studies, new genes and pathways are revealed as causative factors in the development of Parkinson\u2019s disease (PD). However, many of these PD genes are poorly characterized in terms of their function, subcellular localization, and interaction with other components in cellular pathways. This represents a major obstacle towards a better understanding of the molecular causes of PD, with deeper molecular studies often hindered by a lack of high-quality, validated antibodies for detecting the corresponding proteins of interest. In this study, we leveraged the nanoluciferase-derived LgBiT-HiBiT system by generating a cohort of tagged PD genes in both induced pluripotent stem cells (iPSCs) and iPSC-derived neuronal cells. To promote luminescence signals within cells, a master iPSC line was generated, in which LgBiT expression is under the control of a doxycycline-inducible promoter. LgBiT could bind to HiBiT when present either alone or when tagged onto different PD-associated proteins encoded by the genes GBA1, GPNMB, LRRK2, PINK1, PRKN, SNCA, VPS13C, and VPS35. Several HiBiT-tagged proteins could already generate luminescence in iPSCs in response to the doxycycline induction of LgBiT, with the enzyme glucosylceramidase beta 1 (GCase), encoded by GBA1, being one such example. Moreover, the GCase chaperone ambroxol elicited an increase in the luminescence signal in HiBiT-tagged GBA1 cells, correlating with an increase in the levels of GCase in dopaminergic cells. Taken together, we have developed and validated a Doxycycline-inducible luminescence system to serve as a sensitive assay for the quantification, localization, and activity of HiBiT-tagged PD-associated proteins with reliable sensitivity and efficiency.<\/jats:p>","DOI":"10.3390\/ijms25179493","type":"journal-article","created":{"date-parts":[[2024,9,3]],"date-time":"2024-09-03T09:56:42Z","timestamp":1725357402000},"page":"9493","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["An Inducible Luminescent System to Explore Parkinson\u2019s Disease-Associated Genes"],"prefix":"10.3390","volume":"25","author":[{"given":"Anelya","family":"Gandy","sequence":"first","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5785-6238","authenticated-orcid":false,"given":"Gilles","family":"Maussion","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Al-Habyan","sequence":"additional","affiliation":[{"name":"Centre for Oncology, Radiopharmaceuticals and Research (CORR), Biologic and Radiopharmaceutical Drugs Directorate (BRDD), Health Products and Food Branch (HPFB), Health Canada, Ottawa, ON K1A 0K9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Nicouleau","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhipeng","family":"You","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carol X.-Q.","family":"Chen","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Narges","family":"Abdian","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathalia","family":"Aprahamian","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7975-6489","authenticated-orcid":false,"given":"Andrea I.","family":"Krahn","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louise","family":"Larocque","sequence":"additional","affiliation":[{"name":"Centre for Oncology, Radiopharmaceuticals and Research (CORR), Biologic and Radiopharmaceutical Drugs Directorate (BRDD), Health Products and Food Branch (HPFB), Health Canada, Ottawa, ON K1A 0K9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3942-1956","authenticated-orcid":false,"given":"Thomas M.","family":"Durcan","sequence":"additional","affiliation":[{"name":"The Neuro\u2019s Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6875-0380","authenticated-orcid":false,"given":"Eric","family":"Deneault","sequence":"additional","affiliation":[{"name":"Centre for Oncology, Radiopharmaceuticals and Research (CORR), Biologic and Radiopharmaceutical Drugs Directorate (BRDD), Health Products and Food Branch (HPFB), Health Canada, Ottawa, ON K1A 0K9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1111\/j.1460-9568.2009.06873.x","article-title":"The second brain and Parkinson\u2019s disease","volume":"30","author":"Lebouvier","year":"2009","journal-title":"Eur. 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