{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T17:30:18Z","timestamp":1790875818150,"version":"4.1.0"},"reference-count":45,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,2,16]],"date-time":"2023-02-16T00:00:00Z","timestamp":1676505600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Association for Shwachman\u2013Diamond Syndrome","award":["AISS 2018\u2014BALESTRA"],"award-info":[{"award-number":["AISS 2018\u2014BALESTRA"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Shwachman\u2013Diamond syndrome (SDS) represents one of the most common inherited bone marrow failure syndromes and is mainly caused by SBDS gene mutations. Only supportive treatments are available, with hematopoietic cell transplantation required when marrow failure occurs. Among all causative mutations, the SBDS c.258+2T&gt;C variant at the 5\u2032 splice site (ss) of exon 2 is one of the most frequent. Here, we investigated the molecular mechanisms underlying aberrant SBDS splicing and showed that SBDS exon 2 is dense in splicing regulatory elements and cryptic splice sites, complicating proper 5\u2032ss selection. Studies ex vivo and in vitro demonstrated that the mutation alters splicing, but it is also compatible with tiny amounts of correct transcripts, which would explain the survival of SDS patients. Moreover, for the first time for SDS, we explored a panel of correction approaches at the RNA and DNA levels and provided experimental evidence that the mutation effect can be partially counteracted by engineered U1snRNA, trans-splicing, and base\/prime editors, ultimately leading to correctly spliced transcripts (from barely detectable to 2.5\u20135.5%). Among them, we propose DNA editors that, by stably reverting the mutation and potentially conferring positive selection to bone-marrow cells, could lead to the development of an innovative SDS therapy.<\/jats:p>","DOI":"10.3390\/ijms24044024","type":"journal-article","created":{"date-parts":[[2023,2,17]],"date-time":"2023-02-17T01:59:24Z","timestamp":1676599164000},"page":"4024","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Counteracting the Common Shwachman\u2013Diamond Syndrome-Causing SBDS c.258+2T&gt;C Mutation by RNA Therapeutics and Base\/Prime Editing"],"prefix":"10.3390","volume":"24","author":[{"given":"Laura","family":"Peretto","sequence":"first","affiliation":[{"name":"Department of Life Sciences and Biotechnology and LTTA, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Tonetto","sequence":"additional","affiliation":[{"name":"Department of Life Sciences and Biotechnology and LTTA, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iva","family":"Maestri","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and for Romagna, University of Ferrara, 44123 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6849-4487","authenticated-orcid":false,"given":"Valentino","family":"Bezzerri","sequence":"additional","affiliation":[{"name":"Cystic Fibrosis Center, Azienda Ospedaliera Universitaria Integrata di Verona, 37126 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3209-8917","authenticated-orcid":false,"given":"Roberto","family":"Valli","sequence":"additional","affiliation":[{"name":"Department of Medicine and Surgery, University of Insubria, 21100 Varese, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5652-7248","authenticated-orcid":false,"given":"Marco","family":"Cipolli","sequence":"additional","affiliation":[{"name":"Cystic Fibrosis Center, Azienda Ospedaliera Universitaria Integrata di Verona, 37126 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4114-7055","authenticated-orcid":false,"given":"Mirko","family":"Pinotti","sequence":"additional","affiliation":[{"name":"Department of Life Sciences and Biotechnology and LTTA, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6675-9429","authenticated-orcid":false,"given":"Dario","family":"Balestra","sequence":"additional","affiliation":[{"name":"Department of Life Sciences and Biotechnology and LTTA, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1159\/000055858","article-title":"Shwachman-Diamond Syndrome: Clinical Phenotypes","volume":"1","author":"Cipolli","year":"2001","journal-title":"Pancreatology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1038\/ng1062","article-title":"Mutations in SBDS Are Associated with Shwachman-Diamond Syndrome","volume":"33","author":"Boocock","year":"2003","journal-title":"Nat. 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