Scopus
🔓 Açık Erişim YÖKSİS DOI Eşleşti
SJR Q1
Precise CRISPR-Mediated Editing of the TGFBI R555W Mutation in Patient-Derived Peripheral Blood Mononuclear Cells
International Journal of Molecular Sciences · Mart 2026
YÖKSİS Kayıtları
Precise CRISPR-Mediated Editing of the TGFBI R555W Mutation in Patient-Derived Peripheral Blood Mononuclear Cells
International Journal of Molecular Sciences · 2026 SCI-Expanded
Arş. Gör. BURAK DAĞDELEN →
YÖKSİS Kayıtları — ISSN Eşleşmesi
Investigating COX-2 and 5-LOX Enzyme-Related Anti-Inflammatory and Antioxidant Activities and Phytochemical Features of Scutellaria salviifolia Benth
2025 ISSN: 1661-6596 SCI
Arş. Gör. GÜLSÜM METKİN →
Makale Bilgileri
ISSN16616596
Yayın TarihiMart 2026
Cilt / Sayfa27
Scopus ID2-s2.0-105032669792
Erişim🔓 Açık Erişim
Özet
Over 70 mutations in the transforming growth factor beta-induced (TGFBI) gene are associated with corneal dystrophies that impair vision. The R555W hotspot mutation is a major cause of granular corneal dystrophy type 1 (GCD1). Here, we evaluated the technical feasibility of CRISPR/Cas9-mediated editing of the R555W mutation in peripheral blood mononuclear cells (PBMCs) obtained from a patient with GCD1. Three single guide RNAs (sgRNA1–3) and matched single-stranded oligodeoxynucleotide donors (ssODN1–3) were designed and co-transfected into PBMCs. Transfected cells were enriched by flow cytometric sorting, with GFP-positive cells representing approximately 2–4% of the total electroporated population. Editing outcomes were initially screened using high-resolution melting (HRM) analysis, and the sgRNA3–ssODN3 combination identified as the most promising candidate was subsequently validated by next-generation sequencing (NGS). Sequencing revealed a homology-directed repair efficiency of 98.2% among GFP-positive sorted cells, demonstrating efficient and precise genome editing within the enriched population. Because PBMCs are not disease-relevant corneal epithelial cells and only genomic endpoints were assessed, the clinical applicability of this study is limited and the work should be considered a technical proof-of-concept. This framework supports optimization of CRISPR-based strategies prior to studies in biologically relevant corneal models.
Yazarlar (4)
1
Burak Dağdelen
ORCID: 0000-0002-4362-3852
2
Hilal Arikoglu
ORCID: 0000-0002-6600-6603
3
Dudu Erkoc-Kaya
ORCID: 0000-0003-0114-6602
4
Banu Bozkurt
ORCID: 0000-0002-9847-3521
Anahtar Kelimeler
CRISPR/Cas9
genome editing
granular corneal dystrophy
homology-directed repair
peripheral blood mononuclear cells
R555W mutation
TGFBI
Kurumlar
Selçuk Tip Fakültesi
Konya Turkey
Scimago Dergi (ISSN Eşleşmesi)
International Journal of Molecular Sciences
Q1
OA
SJR Skoru1,316
H-Index365
YayıncıMultidisciplinary Digital Publishing Institute (MDPI)
ÜlkeSwitzerland
Catalysis (Q1)
Computer Science Applications (Q1)
Inorganic Chemistry (Q1)
Medicine (miscellaneous) (Q1)
Organic Chemistry (Q1)
Physical and Theoretical Chemistry (Q1)
Spectroscopy (Q1)
Molecular Biology (Q2)