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Cited by 68CRISPR and Genetic EngineeringSkin and Cellular Biology ResearchVirus-based gene therapy research

Ex Vivo COL7A1 Correction for Recessive Dystrophic Epidermolysis Bullosa Using CRISPR/Cas9 and Homology-Directed Repair

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Izmiryan, Araksya, Ganier, Clarisse, Bovolenta, Matteo, Schmitt, Alain, Mavilio, Fulvio, Hovnanian, Alain·2018·Molecular Therapy Nucleic Acids
DOI 10.1016/j.omtn.2018.06.008PMC PMC6077132
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Abstract

Recessive dystrophic epidermolysis bullosa is a rare and severe genetic skin disease resulting in blistering of the skin and mucosa. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by a wide variety of mutations in COL7A1-encoding type VII collagen, which is essential for dermal-epidermal adhesion. Here we demonstrate the feasibility of ex vivo COL7A1 editing in primary RDEB cells and in grafted 3D skin equivalents through CRISPR/Cas9-mediated homology-directed repair. We designed five guide RNAs to correct a RDEB causative null mutation in exon 2 (c.189delG; p.Leu64Trpfs*40). Among the site-specific guide RNAs tested, one showed significant cleavage activity in primary RDEB keratinocytes and in fibroblasts when delivered as integration-deficient lentivirus. Genetic correction was detected in transduced keratinocytes and fibroblasts by allele-specific highly sensitive TaqMan-droplet digital PCR (ddPCR), resulting in 11% and 15.7% of corrected COL7A1 mRNA expression, respectively, without antibiotic selection. Grafting of genetically corrected 3D skin equivalents onto nude mice showed up to 26% re-expression and normal localization of type VII collagen as well as anchoring fibril formation at the dermal-epidermal junction. Our study provides evidence that precise genome editing in primary RDEB cells is a relevant strategy to genetically correct COL7A1 mutations for the development of future ex vivo clinical applications.

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  • CRISPR/Cas9-mediated homology-directed repair via an ex vivo approach conducted gene correction for recessive dystrophic epidermolysis bullosa.

    “Another study also utilised CRISPR/Cas9-mediated, homology-directed repair via an ex vivo approach to conduct gene correction for recessive dystrophic epidermolysis bullosa [120].”

    From Non-Integrating Lentiviral Vectors in Clinical Applications: A Glance Through (2022) · cites this paper as [120]

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Authors
Izmiryan, A., Ganier, C., Bovolenta, M., Schmitt, A., Mavilio, F., & Hovnanian, A.
Affiliations
  • Inserm (FR)
  • University of Modena and Reggio Emilia (IT)
  • Hôpital Necker-Enfants Malades (FR)
Corresponding author
Hovnanian, Alain
ORCID
Venue
Molecular Therapy Nucleic Acids
Publisher
Elsevier BV
Published
2018-09
Volume
12
Pages
554-567
ISSN
2162-2531 (ISSN-L)
DOI
10.1016/j.omtn.2018.06.008
PMC
PMC6077132
OpenAlex
W2811052782
Cited by
68
References
12
Citations by year
2026: 4 · 2025: 5 · 2024: 4 · 2023: 6 · 2022: 12 · 2021: 10 · 2020: 16 · 2019: 10 · 2018: 1
FWCI
3.29
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Therapy — Nucleic Acids (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • IRIS UNIMORE (University of Modena and Reggio Emilia) (submittedVersion, cc-by)
  • HAL (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Institutional Research Information System University of Ferrara (University of Ferrara) (submittedVersion, cc-by)
Funders
  • Debra International
  • Dystrophic Epidermolysis Bullosa Research Association
  • European Research Council (ERC-2010-AdG)
  • Association Française contre les Myopathies
  • Institut National de la Santé et de la Recherche Médicale
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Skin and Cellular Biology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Keywords
recessive dystrophic epidermolysis bullosa, COL7A1, type VII collagen, CRISPR/Cas9, homology-directed repair, ex vivo gene editing, keratinocytes, fibroblasts, 3D skin equivalents, basement membrane zone, anchoring fibrils, lentiviral gene delivery, droplet digital PCR, gene correction

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