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SCA3 patients suffer from ataxia, spasticity and dystonia in mid-adulthood, with spinocerebellar dysfunction and degeneration. As a monogenic disease for which only symptomatic treatment is available, <jats:italic>ATXN3<\/jats:italic> is an attractive target for gene editing. We used the KamiCas9, a self-inactivating gene editing system, to explore gene editing strategies suitable for all SCA3 patients. We first tested the deletion of exon 10 or the introduction of a premature stop codon into exon 9. High editing events were observed in vitro, but efficiency was very low in SCA3 transgenic mice. We then evaluated an ablate-and-replace strategy. The ablate experiments resulted in 55\u2009\u00b1\u200918% cerebellar editing of the <jats:italic>ATXN3<\/jats:italic> gene. A human <jats:italic>ATXN3L<\/jats:italic> paralog, expressed in the brains of SCA3 patients, may act as a natural, CRISPR-resistant replacement gene. In a proof-of-principle study, ablate and ablate-and-replace strategies were evaluated in SCA3 transgenic mice. Two months after injection, similar editing efficiencies were obtained in the ablate and ablate-and-replace groups. Immunofluorescence and RT-qPCR analyses of cerebellar markers support the development of this strategy for SCA3 treatment.<\/jats:p>","DOI":"10.1038\/s41434-025-00557-2","type":"journal-article","created":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T19:25:23Z","timestamp":1753730723000},"page":"462-474","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Gene editing for Spinocerebellar ataxia type 3 taking advantage of the human ATXN3L paralog as replacement gene"],"prefix":"10.1038","volume":"32","author":[{"given":"Margareta","family":"Rybarikova","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Rey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ed","family":"Hasanovic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2955-2629","authenticated-orcid":false,"given":"M\u00e9lanie","family":"Sipion","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lukas","family":"Rambousek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4475-9476","authenticated-orcid":false,"given":"Nicole","family":"D\u00e9glon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,28]]},"reference":[{"key":"557_CR1","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1212\/WNL.22.1.49","volume":"22","author":"KK Nakano","year":"1972","unstructured":"Nakano KK, Dawson DM, Spence A. 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