{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:17:29Z","timestamp":1760059049483,"version":"build-2065373602"},"reference-count":50,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T00:00:00Z","timestamp":1747872000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005762","name":"National Taiwan University Hospital","doi-asserted-by":"publisher","award":["106-T04","107-2622-B-002-008-CC2"],"award-info":[{"award-number":["106-T04","107-2622-B-002-008-CC2"]}],"id":[{"id":"10.13039\/501100005762","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Science and Technology Council of Taiwan","award":["106-T04","107-2622-B-002-008-CC2"],"award-info":[{"award-number":["106-T04","107-2622-B-002-008-CC2"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Recessive variants of SLC26A4 are a common cause of hereditary hearing impairment and are responsible for non-syndromic enlarged vestibular aqueducts and Pendred syndrome. Patients with bi-allelic SLC26A4 variants often suffer from fluctuating hearing loss and recurrent vertigo, ultimately leading to severe to profound hearing impairment. However, there are currently no satisfactory prevention or treatment options for this condition. The CRISPR\/Cas9 genome-editing technique is a well-known tool for correcting point mutations or manipulating genes and shows potential therapeutic applications for hereditary disorders. In this study, we used the homology-independent targeted integration (HITI) strategy to correct the SLC26A4 c.919-2A&gt;G variant, the most common SLC26A4 variant in the Han Chinese population. Next-generation sequencing was performed to evaluate the editing efficiency of the HITI strategy. The results showed that only 0.15% of the reads successfully exhibited HITI integration, indicating that the c.919-2 region may not be a suitable region for HITI selection. This suggests that other site selection or insertion strategies may be needed to improve the efficiency of correcting the SLC26A4 c.919-2A&gt;G variant. This experience may serve as a valuable reference for other researchers considering CRISPR target design in this region.<\/jats:p>","DOI":"10.3390\/ijms26114980","type":"journal-article","created":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T07:04:28Z","timestamp":1747897468000},"page":"4980","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Low Efficiency of Homology-Independent Targeted Integration for CRISPR\/Cas9 Correction in the Vicinity of the SLC26A4 c.919-2A&gt;G Variant"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-3299-6657","authenticated-orcid":false,"given":"Chang-Han","family":"Ho","sequence":"first","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6150-4577","authenticated-orcid":false,"given":"Cheng-Yu","family":"Tsai","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"},{"name":"Graduate Institute of Medical Genomics and Proteomics, National Taiwan University College of Medicine, Taipei 100233, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0783-2868","authenticated-orcid":false,"given":"Chi-Chieh","family":"Chang","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3170-9691","authenticated-orcid":false,"given":"Chin-Ju","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"},{"name":"Program in Speech and Hearing Biosciences and Technology, Harvard Medical School, Boston, MA 02115, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cheng-Yen","family":"Huang","sequence":"additional","affiliation":[{"name":"Gene Knockout\/in Cell Line Modeling Core, Human Disease Modeling Center, First Core Laboratory, Branch Office of Research and Development, National Taiwan University College of Medicine, Taipei 100233, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ying-Chang","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pei-Hsuan","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chin-Hsien","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Neurology, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Han-I","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Neurology, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chih-Hsin","family":"OuYang","sequence":"additional","affiliation":[{"name":"Department of Neurology, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7548-7736","authenticated-orcid":false,"given":"Chuan-Jen","family":"Hsu","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung 427, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6771-5025","authenticated-orcid":false,"given":"Tien-Chen","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6510-3420","authenticated-orcid":false,"given":"You-Tzung","family":"Chen","sequence":"additional","affiliation":[{"name":"Graduate Institute of Medical Genomics and Proteomics, National Taiwan University College of Medicine, Taipei 100233, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yen-Hui","family":"Chan","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"},{"name":"Department of Otolaryngology-Head and Neck Surgery, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung 427, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1995-5854","authenticated-orcid":false,"given":"Yen-Fu","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Medical Research, Taipei Veterans General Hospital, Taipei 112201, Taiwan"},{"name":"Department of Otolaryngology-Head and Neck Surgery, Taipei Veterans General Hospital, Taipei 112201, Taiwan"},{"name":"School of Medicine, National Yang-Ming Chiao-Tung University, Taipei 112304, Taiwan"},{"name":"Department of Speech Language Pathology and Audiology, National Taipei University of Nursing Health Sciences, Taipei 112303, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5047-2204","authenticated-orcid":false,"given":"Chen-Chi","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology-Head and Neck Surgery, National Taiwan University Hospital, Taipei 100225, Taiwan"},{"name":"Department of Medical Research, National Taiwan University Hospital Hsin-Chu Branch, Hsinchu 302058, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.mrrev.2008.08.002","article-title":"Forty-six genes causing nonsyndromic hearing impairment: Which ones should be analyzed in DNA diagnostics?","volume":"681","author":"Hilgert","year":"2009","journal-title":"Mutat. 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