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Trust","award":["SC037861"],"award-info":[{"award-number":["SC037861"]}]},{"name":"LifeArc","award":["5167\/5168"],"award-info":[{"award-number":["5167\/5168"]}]},{"name":"LifeArc","award":["GR001340"],"award-info":[{"award-number":["GR001340"]}]},{"name":"LifeArc","award":["GN3052"],"award-info":[{"award-number":["GN3052"]}]},{"name":"LifeArc","award":["205041\/Z\/16\/Z"],"award-info":[{"award-number":["205041\/Z\/16\/Z"]}]},{"name":"LifeArc","award":["NIHR301696"],"award-info":[{"award-number":["NIHR301696"]}]},{"name":"LifeArc","award":["PGL23\/100048"],"award-info":[{"award-number":["PGL23\/100048"]}]},{"name":"LifeArc","award":["GR001376"],"award-info":[{"award-number":["GR001376"]}]},{"name":"LifeArc","award":["NIHR203312"],"award-info":[{"award-number":["NIHR203312"]}]},{"name":"LifeArc","award":["NIHR203322"],"award-info":[{"award-number":["NIHR203322"]}]},{"name":"LifeArc","award":["10748"],"award-info":[{"award-number":["10748"]}]},{"name":"LifeArc","award":["1015243"],"award-info":[{"award-number":["1015243"]}]},{"name":"LifeArc","award":["SC037861"],"award-info":[{"award-number":["SC037861"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Dominant Optic Atrophy (DOA) is the most common inherited optic neuropathy and presents as gradual visual loss caused by the loss of retinal ganglion cells (RGCs). Over 60% of DOA cases are caused by pathogenic variants in the OPA1 gene, which encodes a mitochondrial GTPase essential in mitochondrial fusion. Currently, there are no treatments for DOA. Here, we tested the therapeutic potential of an approach to DOA using CRISPR activation (CRISPRa). Homology directed repair was used to introduce a common OPA1 pathogenic variant (c.2708_2711TTAGdel) into HEK293T cells as an in vitro model of DOA. Heterozygous c.2708_2711TTAGdel cells had reduced levels of OPA1 mRNA transcript, OPA1 protein, and mitochondrial network alterations. The effect of inactivated Cas9 fused to an activator (dCas9\u2013VPR) was tested with a range of guide RNAs (gRNA) targeted to the promotor region of OPA1. gRNA3 and dCas9\u2013VPR increased OPA1 expression at the RNA and protein level towards control levels. Importantly, the correct ratio of OPA1 isoform transcripts was maintained by CRISPRa. CRISPRa-treated cells showed an improvement in mitochondrial networks compared to untreated cells, indicating partial rescue of a disease-associated phenotype. Collectively, these data support the potential application of CRISPRa as a therapeutic intervention in DOA.<\/jats:p>","DOI":"10.3390\/ijms26136364","type":"journal-article","created":{"date-parts":[[2025,7,2]],"date-time":"2025-07-02T04:11:04Z","timestamp":1751429464000},"page":"6364","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["CRISPRa-Mediated Increase of OPA1 Expression in Dominant Optic Atrophy"],"prefix":"10.3390","volume":"26","author":[{"given":"Giada","family":"Becchi","sequence":"first","affiliation":[{"name":"UCL Institute of Ophthalmology, Faculty of Brain Sciences, London EC1V 9EL, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Whitehead","sequence":"additional","affiliation":[{"name":"UCL Institute of Ophthalmology, Faculty of Brain Sciences, London EC1V 9EL, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joshua P.","family":"Harvey","sequence":"additional","affiliation":[{"name":"UCL Institute of Ophthalmology, Faculty of Brain Sciences, London EC1V 9EL, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul E.","family":"Sladen","sequence":"additional","affiliation":[{"name":"UCL Institute of Ophthalmology, Faculty of Brain Sciences, London EC1V 9EL, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohammed","family":"Dushti","sequence":"additional","affiliation":[{"name":"William Harvey Research Institute, Faculty of Medicine and Dentistry, Queen Mary University of London, London EC1M 6BQ, UK"},{"name":"Pharmacology & Toxicology Department, Faculty of Medicine, Kuwait University, Kuwait City P.O. Box 24923, Kuwait"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1876-1505","authenticated-orcid":false,"given":"J. Paul","family":"Chapple","sequence":"additional","affiliation":[{"name":"William Harvey Research Institute, Faculty of Medicine and Dentistry, Queen Mary University of London, London EC1M 6BQ, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7847-9320","authenticated-orcid":false,"given":"Patrick","family":"Yu-Wai-Man","sequence":"additional","affiliation":[{"name":"John van Geest Centre for Brain Repair, University of Cambridge, Cambridge CB2 0PY, UK"},{"name":"MRC Mitochondrial Biology Unit, Department of Clinical Neurosciences, University of Cambridge, Cambridge CB2 0XY, UK"},{"name":"Cambridge Eye Unit, Addenbrooke\u2019s Hospital, Cambridge University Hospital NHS Foundation Trust, Cambridge CB2 0QQ, UK"},{"name":"Moorfields Eye Hospital NHS Foundation Trust, London EC1V 2PD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6429-654X","authenticated-orcid":false,"given":"Michael E.","family":"Cheetham","sequence":"additional","affiliation":[{"name":"UCL Institute of Ophthalmology, Faculty of Brain Sciences, London EC1V 9EL, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/j.1600-0420.1996.tb00672.x","article-title":"Dominant Optic Atrophy Mapped to Chromosome 3q Region: II. 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Ther.-Nucleic Acids"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/13\/6364\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:02:46Z","timestamp":1760032966000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/13\/6364"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,2]]},"references-count":45,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["ijms26136364"],"URL":"https:\/\/doi.org\/10.3390\/ijms26136364","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,2]]}}}