CRISPR/Cas9-Mediated Treatment Ameliorates the Phenotype of the Epidermolytic Palmoplantar Keratoderma-like Mouse
| Autor: | Xiang Gao, Hai-Ping Ke, Xiao-Rui Luan, Zhaoyu Lin, Yuexiao Tang, Xiao-Ling Chen, J. Zhang, Xian-Ning Zhang |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Genetic enhancement Mutant Biology medicine.disease_cause Keratin disease Article 03 medical and health sciences 0302 clinical medicine epidermolytic palmoplantar keratoderma Drug Discovery medicine CRISPR CRISPR/Cas9 Mutation Epidermolytic Palmoplantar Keratoderma Cas9 lcsh:RM1-950 Krt9 gene (mouse) medicine.disease Molecular biology gene therapy Protospacer adjacent motif lcsh:Therapeutics. Pharmacology 030104 developmental biology indel 030220 oncology & carcinogenesis Molecular Medicine |
| Zdroj: | Molecular Therapy. Nucleic Acids Molecular Therapy: Nucleic Acids, Vol 12, Iss, Pp 220-228 (2018) |
| ISSN: | 2162-2531 |
| Popis: | CRISPR/Cas9 has been confirmed as a distinctly efficient, simple-to-configure, highly specific genome-editing tool that has been used to treat monogenetic disorders. Epidermolytic palmoplantar keratoderma (EPPK) is a common autosomal dominant keratin disease resulting from dominant-negative mutation of the KRT9 gene, and it has no effective therapy. We performed CRISPR/Cas9-mediated treatment on a knockin (KI) transgenic mouse model that carried a small indel heterozygous mutation of Krt9, c.434delAinsGGCT (p.Tyr144delinsTrpLeu), which caused a humanized EPPK-like phenotype. The mutation within exon 1 of Krt9 generated a novel protospacer adjacent motif site, TGG, for Cas9 recognition and cutting. By delivering lentivirus vectors (LVs) encoding single-guide RNAs (sgRNAs) and Cas9 that targeted Krt9 sequence into HeLa cells engineered to constitutively express wild-type and mutant keratin 9 (K9), we found the sgRNA was highly effective in reducing expression of the mutant K9 protein in vitro. We injected the LV into the fore-paws of adult KI-Krt9 mice three times every 8 days and found that the expression of K9 decreased ∼14.6%. The phenotypic mitigation was revealed by restoration of the abnormal differentiation and aberrant proliferation of the epidermis. Our data are the first to show that CRISPR/Cas9 is a potentially powerful therapeutic option for EPPK and other PPK subtypes. Keywords: CRISPR/Cas9, gene therapy, epidermolytic palmoplantar keratoderma, Krt9 gene (mouse), indel |
| Databáze: | OpenAIRE |
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