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Background: Recent evidence suggests a crucial biological role for Circular RNAs (circRNAs) in keloid diseases, yet the underlying mechanisms remain unclear. This study explored the biological effects and molecular mechanisms of hsa_circ_0002198 in keloid formation. Methods: Real-time quantitative PCR (qRT-PCR) was employed to assess the expression of circ_0002198 in keloid tissues, normal skin tissues, keloid fibroblasts (KFs), and normal skin fibroblasts (NFs) from nine patients. To investigate the role of circ_0002198 in keloid pathogenesis, cell transfection technology was utilized to knock down circ_0002198. Various experiments including Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2′-deoxyuridine (EdU), Transwell, wound healing assay, flow cytometry, and others were conducted to explore the potential mechanisms associated with circ_0002198 expression. The RNA-binding protein Eukaryotic translation initiation factor 4A, isoform 3 (EIF4A3) binding to circ_0002198 was identified and confirmed through bioinformatics databases prediction and RNA immunoprecipitation (RIP) assay. Finally, the expression of EIF4A3 was assessed, and both silencing and overexpression were employed to verify its role in circ_0002198 regulation. Results: The expression levels of circ_0002198 and EIF4A3 were notably elevated in keloid tissues and KFs compared to normal skin tissues and NFs. The reduction of circ_0002198 expression in KFs significantly impeded their proliferation, migration, and invasion. It also hindered the cell cycle process and the expression of associated proteins while concurrently promoting apoptosis in KFs. EIF4A3 was identified to bind to the flanks of circ_0002198, enhancing the occurrence of circ_0002198 and its role in regulating the progression of KFs. Conclusion: Our study offers insights into how Circular RNA may contribute to the pathogenesis of keloid formation, highlighting Circ_0002198 as a potential novel biomarker for keloids in association with EIF4A3. Further research, involving larger study cohorts, is necessary to broaden our understanding of keloid mechanisms and potential treatment approaches. [ABSTRACT FROM AUTHOR]

• We discuss the advantages and disadvantages of AAV as vectors for gene therapy. • We describe the methods for AAV optimization. • The challenges and solutions for delivering CRISPR/Cas9 system using AAV as a vectors in gene therapy have been shown in this article. The adeno-associated virus (AAV) is a defective single-stranded DNA virus with the simplest structure reported to date. It constitutes a capsid protein and single-stranded DNA. With its high transduction efficiency, low immunogenicity, and tissue specificity, it is the most widely used and promising gene therapy vector. The clustered regularly interspaced short palindromic sequence (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing system is an emerging technology that utilizes cas9 nuclease to specifically recognize and cleave target genes under the guidance of small guide RNA and realizes gene editing through homologous directional repair and non-homologous recombination repair. In recent years, an increasing number of animal experiments and clinical studies have revealed the great potential of AAV as a vector to deliver the CRISPR/cas9 system for treating genetic diseases and viral infections. However, the immunogenicity, toxicity, low transmission efficiency in brain and ear tissues, packaging size limitations of AAV, and immunogenicity and off-target effects of Cas9 protein pose several clinical challenges. This research reviews the role, challenges, and countermeasures of the AAV-CRISPR/cas9 system in gene therapy. [ABSTRACT FROM AUTHOR]