{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T22:00:07Z","timestamp":1790460007906,"version":"4.1.0"},"reference-count":50,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2020,11,20]],"date-time":"2020-11-20T00:00:00Z","timestamp":1605830400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100009873","name":"Regione Autonoma della Sardegna","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009873","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Br J Haematol"],"published-print":{"date-parts":[[2021,1]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Hereditary persistence of fetal haemoglobin (HPFH) is the major modifier of the clinical severity of \u03b2\u2010thalassaemia. The homozygous mutation c.\u2010196 C&gt;T in the A\u03b3\u2010globin (<jats:italic>HBG1<\/jats:italic>) promoter, which causes Sardinian \u03b4\u03b2<jats:sup>0<\/jats:sup>\u2010thalassaemia, is able to completely rescue the \u03b2\u2010major thalassaemia phenotype caused by the \u03b2<jats:sup>0<\/jats:sup>39\u2010thalassaemia mutation, ensuring high levels of fetal haemoglobin synthesis during adulthood. Here, we describe a CRISPR\/Cas9 genome\u2010editing approach, combined with the non\u2010homologous end joining (NHEJ) pathway repair, aimed at reproducing the effects of this naturally occurring HPFH mutation in both <jats:italic>HBG<\/jats:italic> promoters. After selecting the most efficient guide RNA in K562 cells, we edited the <jats:italic>HBG<\/jats:italic> promoters in human umbilical cord blood\u2010derived erythroid progenitor 2 cells (HUDEP\u20102) and in haematopoietic stem and progenitor cells (HSPCs) from \u03b2<jats:sup>0<\/jats:sup>\u2010thalassaemia patients to assess the therapeutic potential of HbF induction. Our results indicate that small deletions targeting the \u2212196\u2010promoter region restore high levels of fetal haemoglobin (HbF) synthesis in all cell types tested. In pools of HSPCs derived from homozygous \u03b2<jats:sup>0<\/jats:sup>39\u2010thalassaemia patients, a 20% editing determined a parallel 20% increase of HbF compared to unedited pools. These results suggest that editing the region of <jats:italic>HBG<\/jats:italic> promoters around the \u2212196 position has the potential to induce therapeutic levels of HbF in patients with most types of \u03b2\u2010thalassaemia irrespective of the \u03b2\u2010globin gene (<jats:italic>HBB<\/jats:italic>) mutations.<\/jats:p>","DOI":"10.1111\/bjh.17167","type":"journal-article","created":{"date-parts":[[2020,11,21]],"date-time":"2020-11-21T06:21:05Z","timestamp":1605939665000},"page":"395-404","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Induction of therapeutic levels of HbF in genome\u2010edited primary \u03b2<sup>0<\/sup>39\u2010thalassaemia haematopoietic stem and progenitor cells"],"prefix":"10.1111","volume":"192","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3716-241X","authenticated-orcid":false,"given":"Maura","family":"Mingoia","sequence":"first","affiliation":[{"name":"Dipartimento di Scienze Mediche e Sanit\u00e0 Pubblica Universit\u00e0 degli Studi di Cagliari Cagliari 09124 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6806-3492","authenticated-orcid":false,"given":"Cristian A.","family":"Caria","sequence":"additional","affiliation":[{"name":"Istituto di Ricerca Genetica e Biomedica (IRGB) Consiglio Nazionale Ricerche Monserrato 09042 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7351-8685","authenticated-orcid":false,"given":"Lin","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Medicine and Institute for Human Genetics University of California San Francisco CA 93143 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4389-2470","authenticated-orcid":false,"given":"Isadora","family":"Asunis","sequence":"additional","affiliation":[{"name":"Istituto di Ricerca Genetica e Biomedica (IRGB) Consiglio Nazionale Ricerche Monserrato 09042 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3363-8046","authenticated-orcid":false,"given":"M. Franca","family":"Marongiu","sequence":"additional","affiliation":[{"name":"Istituto di Ricerca Genetica e Biomedica (IRGB) Consiglio Nazionale Ricerche Monserrato 09042 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1800-5139","authenticated-orcid":false,"given":"Laura","family":"Manunza","sequence":"additional","affiliation":[{"name":"Dipartimento di Scienze Mediche e Sanit\u00e0 Pubblica Universit\u00e0 degli Studi di Cagliari Cagliari 09124 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4228-2152","authenticated-orcid":false,"given":"M. Carla","family":"Sollaino","sequence":"additional","affiliation":[{"name":"Ospedale Pediatrico \u201cA. Cao\u201d \u2013 A.O. \u201cG. Brotzu\u201d Cagliari 09121 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5504-4276","authenticated-orcid":false,"given":"Jiaming","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Medicine and Institute for Human Genetics University of California San Francisco CA 93143 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8216-2120","authenticated-orcid":false,"given":"Annalisa","family":"Cabriolu","sequence":"additional","affiliation":[{"name":"Center for Cell Engineering Memorial Sloan Kettering Cancer Centre New York NY 10065 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryo","family":"Kurita","sequence":"additional","affiliation":[{"name":"Cell Engineering Division RIKEN BioResource Center Tsukuba Ibaraki 305\u20100074 Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukio","family":"Nakamura","sequence":"additional","affiliation":[{"name":"Cell Engineering Division RIKEN BioResource Center Tsukuba Ibaraki 305\u20100074 Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7414-1995","authenticated-orcid":false,"given":"Francesco","family":"Cucca","sequence":"additional","affiliation":[{"name":"Istituto di Ricerca Genetica e Biomedica (IRGB) Consiglio Nazionale Ricerche Monserrato 09042 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0117-0100","authenticated-orcid":false,"given":"Yuet W.","family":"Kan","sequence":"additional","affiliation":[{"name":"Department of Medicine and Institute for Human Genetics University of California San Francisco CA 93143 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2359-2892","authenticated-orcid":false,"given":"M. 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