CRISPR-Cas9 Applications in Gene Therapy: Advances, Challenges, and Future Perspectives

Abstract

CRISPR-Cas9 has rapidly emerged as a gold standard for accurate genome editing, which boasts remarkable potential for human therapies. This brief review article summarizes the most up-to-date developments in CRISPR-Cas9 gene therapy with a focus on single-gene disorders such as sickle cell disease, β-thalassemia, and Leber congenital amaurosis. We explain how Cas9 generates a double-stranded break in DNA and how subsequent repair leads to gene correction. We describe base and prime editing methods that raise precision and drastically reduce off-target effects. Still, one of the most significant challenges in the therapeutic context is the delivery of the editing machinery to the desired cells. There are many approaches under investigation, such as the use of adeno-associated viral vectors, lipid nanoparticles or electroporation, which are being evaluated for their efficacy, safety, and their ability to home in on the right tissue. Immunogenicity, undesired mutations, and long-term genetic stability remain major concerns. Besides the technical concerns, we also go over the ethical and legal issues, such as the germline gene editing, equitable access, and informed consent, which emphasize the whole world perspective on responsible use. Eventually, the therapeutic scope of CRISPR-Cas9 may be expanded by joining forces with RNA-targeting technologies, epigenetic modulators, and AI-based design software. Despite the hurdles, CRISPR-Cas9 is expected to revolutionize the field of precision medicine, thereby providing extraordinary possibilities for safe and efficacious genome-based medical interventions.