APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel Variants Article Swipe
YOU?
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· 2022
· Open Access
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· DOI: https://doi.org/10.3390/ijms23084281
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.
Related Topics
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3390/ijms23084281
- https://www.mdpi.com/1422-0067/23/8/4281/pdf?version=1649829661
- OA Status
- gold
- Cited By
- 10
- References
- 58
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4224102582
Raw OpenAlex JSON
- OpenAlex ID
-
https://openalex.org/W4224102582Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.3390/ijms23084281Digital Object Identifier
- Title
-
APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel VariantsWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2022Year of publication
- Publication date
-
2022-04-13Full publication date if available
- Authors
-
Xavier Vanhoye, Alexandre Janin, Amandine Caillaud, Antoine Rimbert, Fabienne Venet, Morgane Gossez, Wieneke Dijk, Oriane Marmontel, Séverine Nony, C. Chatelain, Christine Durand, Pierre Lindenbaum, Jennifer Rieusset, Bertrand Cariou, Philippe Moulin, Mathilde Di FilippoList of authors in order
- Landing page
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https://doi.org/10.3390/ijms23084281Publisher landing page
- PDF URL
-
https://www.mdpi.com/1422-0067/23/8/4281/pdf?version=1649829661Direct link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
- OA URL
-
https://www.mdpi.com/1422-0067/23/8/4281/pdf?version=1649829661Direct OA link when available
- Concepts
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CRISPR, Computational biology, Apolipoprotein B, Biology, Genetics, Gene, Biochemistry, CholesterolTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
10Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 2, 2024: 1, 2023: 6, 2022: 1Per-year citation counts (last 5 years)
- References (count)
-
58Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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