{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T16:39:26Z","timestamp":1788367166424,"version":"build-2803163510"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,4,13]],"date-time":"2022-04-13T00:00:00Z","timestamp":1649808000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006451","name":"Hospices Civils de Lyon","doi-asserted-by":"publisher","award":["Young Researcher Project"],"award-info":[{"award-number":["Young Researcher Project"]}],"id":[{"id":"10.13039\/501100006451","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR-16-RHUS-0007"],"award-info":[{"award-number":["ANR-16-RHUS-0007"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Fondation Recherche M\u00e9dicale","award":["Fondation Recherche M\u00e9dicale"],"award-info":[{"award-number":["Fondation Recherche M\u00e9dicale"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hypobetalipoproteinemia is characterized by LDL-cholesterol and apolipoprotein B (apoB) plasma levels below the fifth percentile for age and sex. Familial hypobetalipoproteinemia (FHBL) is mostly caused by premature termination codons in the APOB gene, a condition associated with fatty liver and steatohepatitis. Nevertheless, many families with a FHBL phenotype carry APOB missense variants of uncertain significance (VUS). We here aimed to develop a proof-of-principle experiment to assess the pathogenicity of VUS using the genome editing of human liver cells. We identified a novel heterozygous APOB-VUS (p.Leu351Arg), in a FHBL family. We generated APOB knock-out (KO) and APOB-p.Leu351Arg knock-in Huh7 cells using CRISPR-Cas9 technology and studied the APOB expression, synthesis and secretion by digital droplet PCR and ELISA quantification. The APOB expression was decreased by 70% in the heterozygous APOB-KO cells and almost abolished in the homozygous-KO cells, with a consistent decrease in apoB production and secretion. The APOB-p.Leu351Arg homozygous cells presented with a 40% decreased APOB expression and undetectable apoB levels in cellular extracts and supernatant. Thus, the p.Leu351Arg affected the apoB secretion, which led us to classify this new variant as likely pathogenic and to set up a hepatic follow-up in this family. Therefore, the functional assessment of APOB-missense variants, using gene-editing technologies, will lead to improvements in the molecular diagnosis of FHBL and the personalized follow-up of these patients.<\/jats:p>","DOI":"10.3390\/ijms23084281","type":"journal-article","created":{"date-parts":[[2022,4,13]],"date-time":"2022-04-13T21:33:42Z","timestamp":1649885622000},"page":"4281","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel Variants"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8875-5851","authenticated-orcid":false,"given":"Xavier","family":"Vanhoye","sequence":"first","affiliation":[{"name":"Service de Biochimie et de Biologie Mol\u00e9culaire, Laboratoire de Biologie M\u00e9dicale MultiSites, Hospices Civils de Lyon, F-69677 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Janin","sequence":"additional","affiliation":[{"name":"Service de Biochimie et de Biologie Mol\u00e9culaire, Laboratoire de Biologie M\u00e9dicale MultiSites, Hospices Civils de Lyon, F-69677 Bron, France"},{"name":"Institut NeuroMyoG\u00e8ne, CNRS UMR5310, INSERM U1217, Universit\u00e9 Claude Bernard Lyon 1, Universit\u00e9 de Lyon, F-69008 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amandine","family":"Caillaud","sequence":"additional","affiliation":[{"name":"Institut du Thorax, Nantes Universit\u00e9, CHU Nantes, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2971-4926","authenticated-orcid":false,"given":"Antoine","family":"Rimbert","sequence":"additional","affiliation":[{"name":"Institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabienne","family":"Venet","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie, Edouard Herriot Hospital, Hospices Civils de Lyon, F-69437 Lyon, France"},{"name":"Centre International de Recherche en Infectiologie (CIRI), INSERM U1111, CNRS, UMR5308, Ecole Normale Sup\u00e9rieure de Lyon, Universit\u00e9 Claude Bernard-Lyon 1, F-69364 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1930-8956","authenticated-orcid":false,"given":"Morgane","family":"Gossez","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie, Edouard Herriot Hospital, Hospices Civils de Lyon, F-69437 Lyon, France"},{"name":"Centre International de Recherche en Infectiologie (CIRI), INSERM U1111, CNRS, UMR5308, Ecole Normale Sup\u00e9rieure de Lyon, Universit\u00e9 Claude Bernard-Lyon 1, F-69364 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1676-7751","authenticated-orcid":false,"given":"Wieneke","family":"Dijk","sequence":"additional","affiliation":[{"name":"Institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oriane","family":"Marmontel","sequence":"additional","affiliation":[{"name":"Service de Biochimie et de Biologie Mol\u00e9culaire, Laboratoire de Biologie M\u00e9dicale MultiSites, Hospices Civils de Lyon, F-69677 Bron, France"},{"name":"CarMen Laboratory, INSERM, INRA, INSA Lyon, Universit\u00e9 Claude Bernard Lyon 1, Pierre-B\u00e9nite, F-69364 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00e9verine","family":"Nony","sequence":"additional","affiliation":[{"name":"Service de Biochimie et de Biologie Mol\u00e9culaire, Laboratoire de Biologie M\u00e9dicale MultiSites, Hospices Civils de Lyon, F-69677 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlotte","family":"Chatelain","sequence":"additional","affiliation":[{"name":"Service de Biochimie et de Biologie Mol\u00e9culaire, Laboratoire de Biologie M\u00e9dicale MultiSites, Hospices Civils de Lyon, F-69677 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"Durand","sequence":"additional","affiliation":[{"name":"CarMen Laboratory, INSERM, INRA, INSA Lyon, Universit\u00e9 Claude Bernard Lyon 1, Pierre-B\u00e9nite, F-69364 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0148-9787","authenticated-orcid":false,"given":"Pierre","family":"Lindenbaum","sequence":"additional","affiliation":[{"name":"Institut du Thorax, Nantes Universit\u00e9, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Rieusset","sequence":"additional","affiliation":[{"name":"CarMen Laboratory, INSERM, INRA, INSA Lyon, Universit\u00e9 Claude Bernard Lyon 1, Pierre-B\u00e9nite, F-69364 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bertrand","family":"Cariou","sequence":"additional","affiliation":[{"name":"Institut du Thorax, Nantes Universit\u00e9, CHU Nantes, CNRS, INSERM, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philippe","family":"Moulin","sequence":"additional","affiliation":[{"name":"CarMen Laboratory, INSERM, INRA, INSA Lyon, Universit\u00e9 Claude Bernard Lyon 1, Pierre-B\u00e9nite, F-69364 Lyon, France"},{"name":"F\u00e9d\u00e9ration d\u2019Endocrinologie, Maladies M\u00e9taboliques, Diab\u00e8te et Nutrition, H\u00f4pital Louis Pradel, Hospices Civils de Lyon, F-69677 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6206-5908","authenticated-orcid":false,"given":"Mathilde","family":"Di Filippo","sequence":"additional","affiliation":[{"name":"Service de Biochimie et de Biologie Mol\u00e9culaire, Laboratoire de Biologie M\u00e9dicale MultiSites, Hospices Civils de Lyon, F-69677 Bron, France"},{"name":"CarMen Laboratory, INSERM, INRA, INSA Lyon, Universit\u00e9 Claude Bernard Lyon 1, Pierre-B\u00e9nite, F-69364 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,4,13]]},"reference":[{"key":"ref_1","unstructured":"Scriver, C., Beaudet, A., Valle, D., and Sly, W. 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