{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T11:35:56Z","timestamp":1790681756881,"version":"4.1.0"},"reference-count":64,"publisher":"American Society for Clinical Investigation","issue":"15","funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R01- HL 064274"],"award-info":[{"award-number":["R01- HL 064274"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2019,8,8]]},"abstract":"<jats:p>\n                    Nonintegrative AAV-mediated gene therapy in the liver is effective in adult patients but faces limitations in pediatric settings because of episomal DNA loss during hepatocyte proliferation. Gene targeting is a promising approach as it results in the permanent modification of the genome. We previously rescued neonatal lethality in Crigler-Najjar mice by inserting a promoterless human uridine glucuronosyl transferase A1 (\n                    <jats:italic>UGT1A1<\/jats:italic>\n                    ) cDNA in exon 14 of the albumin gene, without the use of nucleases. To increase the recombination rate and therapeutic efficacy, we used CRISPR\/\n                    <jats:italic>Sa<\/jats:italic>\n                    Cas9. Neonatal mice were transduced with 2 AAVs: one expressing the\n                    <jats:italic>Sa<\/jats:italic>\n                    Cas9 and sgRNA and one containing a promoterless cDNA flanked by albumin homology regions. Targeting efficiency increased approximately 26-fold with an\n                    <jats:italic>EGFP<\/jats:italic>\n                    reporter cDNA, reaching up to 24% of EGFP-positive hepatocytes. Next, we fully corrected the diseased phenotype of Crigler-Najjar mice by targeting the\n                    <jats:italic>hUGT1A1<\/jats:italic>\n                    cDNA. Treated mice had normal plasma bilirubin up to 10 months after administration, hUGT1A1 protein levels were approximately 6-fold higher than in WT liver, with a 90-fold increase in recombination rate. Liver histology, inflammatory markers, and plasma albumin were normal in treated mice, with no off-targets in predicted sites. Thus, the improved efficacy and reassuring safety profile support the potential application of the proposed approach to other liver diseases.\n                  <\/jats:p>","DOI":"10.1172\/jci.insight.128863","type":"journal-article","created":{"date-parts":[[2019,6,18]],"date-time":"2019-06-18T12:01:43Z","timestamp":1560859303000},"source":"Crossref","is-referenced-by-count":41,"title":["Coupling AAV-mediated promoterless gene targeting to SaCas9 nuclease to efficiently correct liver metabolic diseases"],"prefix":"10.1172","volume":"4","author":[{"given":"Alessia","family":"De Caneva","sequence":"first","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabiola","family":"Porro","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, 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