{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T08:58:04Z","timestamp":1790499484734,"version":"4.1.0"},"reference-count":59,"publisher":"SAGE Publications","issue":"9","license":[{"start":{"date-parts":[[2019,9,1]],"date-time":"2019-09-01T00:00:00Z","timestamp":1567296000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Human Gene Therapy"],"published-print":{"date-parts":[[2019,9]]},"abstract":"<jats:p>\n                    Early efforts in cystic fibrosis (CF) gene therapy faced major challenges in delivery efficiency and sustained therapeutic gene expression. Recent advancements in engineered site-specific endonucleases such as clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas9 make permanent CF transmembrane conductance regulator (\n                    <jats:italic toggle=\"yes\">CFTR<\/jats:italic>\n                    ) gene correction possible. However, because of safety concerns of the CRISPR\/Cas9 system and challenges in\n                    <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                    delivery to inflamed CF airway, CRISPR-based gene correction strategies need to be tested in proper animal models. In this study, we aimed at creating vectors for testing\n                    <jats:italic toggle=\"yes\">CFTR<\/jats:italic>\n                    gene correction in pig models. We constructed helper-dependent adenoviral (HD-Ad) vectors to deliver CRISPR\/Cas9 and a donor template (a 6\u2009kb\n                    <jats:italic toggle=\"yes\">LacZ<\/jats:italic>\n                    or 8.7\u2009kb human\n                    <jats:italic toggle=\"yes\">CFTR<\/jats:italic>\n                    expression cassette) into cultured pig cells. We demonstrated precise integration of each donor into the\n                    <jats:italic toggle=\"yes\">GGTA1<\/jats:italic>\n                    safe harbor through Cas9-induced homology directed repair with 3\u2009kb homology arms. In addition, we showed that both\n                    <jats:italic toggle=\"yes\">LacZ<\/jats:italic>\n                    and\n                    <jats:italic toggle=\"yes\">hCFTR<\/jats:italic>\n                    were persistently expressed in transduced cells. Furthermore, we created a CFTR-deficient cell line for testing CFTR correction. We detected hCFTR mRNA and protein expression in cells transduced with the hCFTR vector. We also demonstrated CFTR function in the CF cells transduced with the HD-Ad delivering the CRISPR-Cas9 system and hCFTR donor at late cellular passages using the membrane potential sensitive dye-based assay (FLIPR\n                    <jats:sup>\u00ae<\/jats:sup>\n                    ). Combined with our previous report on gene delivery to pig airway basal cells, these data provide the feasibility of testing CRISPR\/Cas9-mediated permanent human CFTR correction through HD-Ad vector delivery in pigs.\n                  <\/jats:p>","DOI":"10.1089\/hum.2019.074","type":"journal-article","created":{"date-parts":[[2019,5,17]],"date-time":"2019-05-17T05:14:30Z","timestamp":1558070070000},"page":"1101-1116","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":37,"title":["<i>In Vitro<\/i>\n                    Validation of a CRISPR-Mediated CFTR Correction Strategy for Preclinical Translation in Pigs"],"prefix":"10.1177","volume":"30","author":[{"given":"Zhichang Peter","family":"Zhou","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada"},{"name":"Program of Translational Medicine, The Hospital for Sick Children, Toronto, Canada"},{"name":"Zebrafish Centre for Advanced Drug Discovery and Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Toronto, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang Leo","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada"},{"name":"Program of Translational Medicine, The Hospital for Sick Children, Toronto, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huibi","family":"Cao","sequence":"additional","affiliation":[{"name":"Program of Translational Medicine, The Hospital for Sick Children, Toronto, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyan Rachel","family":"Chen","sequence":"additional","affiliation":[{"name":"Program of Translational Medicine, The Hospital for Sick Children, Toronto, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiqian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Program of Translational Medicine, The Hospital for Sick Children, Toronto, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao-Yan","family":"Wen","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada"},{"name":"Zebrafish Centre for Advanced Drug Discovery and Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Toronto, Canada"},{"name":"Department of Medicine, Physiology and Institute of Medical Science, University of Toronto, Toronto, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jim","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada"},{"name":"Program of Translational Medicine, The 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