Aug 2026· International Journal of Basic & Clinical Pharmacology· Vol 15, pp. 1089-1094· 0 citations· 17 references
TL;DR
Clinical applicability is limited by issues such off-target effects, PAM sequence restrictions, DNA damage-induced toxicity, and immunological responses to Cas proteins, despite its wide therapeutic potential, but improvements in delivery methods and high-fidelity Cas9 variations are being addressed.
Abstract
The adaptive immune system of prokaryotes is the source of CRISPR-Cas9, a ground-breaking genome editing technique that uses RNA-guided nucleases to precisely alter DNA sequences. An summary of CRISPR/Cas9's discovery, structural elements, mode of action, therapeutic uses, and present limitations is given in this article. The Cas9 nuclease and guide RNA work together to identify particular DNA targets and cause double-strand breaks in the system. Cellular processes like homology-directed repair or non-homologous end joining fix these defects, allowing for gene disruption or correction. Numerous genetic abnormalities, such as hemoglobinopathies including sickle cell disease and β-thalassemia, hereditary retinal diseases, muscular dystrophies, liver metabolic disorders, congenital lung diseases, and genetic deafness, have showed great potential for treatment with CRISPR/Cas9. Clinical applicability is limited by issues such off-target effects, PAM sequence restrictions, DNA damage-induced toxicity, and immunological responses to Cas proteins, despite its wide therapeutic potential. These obstacles are being addressed by improvements in delivery methods and high-fidelity Cas9 variations. All things considered, CRISPR/Cas9 is a revolutionary development in molecular medicine and gene therapy, providing strong prospects for accurate genome engineering and upcoming clinical uses in personalized medicine.
This review systematically summarizes the developmental logic, core mechanisms, clinical applications, advantages and limitations of the three generations of CRISPR-Cas technology in monogenic disorders, and analyzes the key challenges such as delivery efficiency, long-term safety, and treatment accessibility.
Yiwen Wang· Theoretical and Natural Scie...· 0 citations
This article synthesizes contemporary advancements in CRISPR-mediated mammalian genome modification, detailing core mechanisms – such as guide RNA and the Cas9 endonuclease – alongside next-generation modalities, including base and prime editing.
Olga Aldoshina, Dmitriy Lazarev, E. Smirnova· Veterinariya, Zootekhniya i...· 0 citations
This chapter outlines a comprehensive methodology for the design, assembly, and functional assessment of CRISPR/dCas9 systems optimized for tomato to investigate pathogen-associated responses.
Ananya Mukherjee, Shrabani Basak, Raghuvir Singh et al.· Methods in molecular biology· 0 citations
CRISPR has emerged as a next-generation gene-editing tool with the potential to target the molecular pathways associated with ageing and related disorders. It functions through RNA-guided Cas nucleases, directing DNA cleavage and utilizing the native DNA repair machinery for genetic manipulations. Advances in CRISPR technology have significantly enhanced the precision and flexibility of techniques for genome editing. The enzyme Cas9's ability to cut DNA at exact site has revolutionized genome editing by enabling accurate modifications within living eukaryotic cells. This review critically examines recent developments in CRISPR-based technologies, including Cas9, Cas12, base editing, prime editing, and CRISPR-mediated gene regulation. It highlights their rising applications in ageing research, with more emphasis on neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. The review also discusses the major pharmacological and translational challenges that currently limit clinical applications, including inefficient tissue-specific delivery, off-target genome editing, immunogenicity, manufacturing complexity, and long-term safety concerns. Also, recent progress in both, viral and non-viral delivery methods are critically evaluated, including adeno-associated viruses, lentivirus vectors, lipid nanoparticles, gold nanoparticles, exosomes, electroporation, and microinjection, is thoroughly discussed to highlight their therapeutic potential and translational limitations. Current studies indicate that CRISPR-based approaches have preclinical potential for targeting important hallmarks of ageing, particularly genomic instability, telomere attrition, and mitochondrial dysfunction. Other hallmarks of ageing, such as stem cell exhaustion, epigenetic modifications, and microbiome changes, are at earlier stages of development. Overall, this review describes future strategies for developing safe, precise, and clinically translatable CRISPR-based treatments to promote healthy ageing.
Sakshi Rathore, Akash Gupta, Kamal Shah et al.· Ageing Research Reviews· 0 citations
Examination of the predicted secondary structure of the tracrRNA–crRNA duplex suggests that the features required for Cas9-catalyzed DNA cleavage at specific sites can be captured within a single chimeric RNA.
A. Udristioiu, Manole Cojocaru· Clinical Cancer Research· 0 citations
Examination of the predicted secondary structure of the tracrRNA–crRNA duplex suggests that the features required for Cas9-catalyzed DNA cleavage at specific sites can be captured within a single chimeric RNA.
A. Udristioiu, Manole Cojocaru· Cancer Research· 0 citations
