Long-read sequencing and next-generation CRISPR editors: a unified pipeline for rare disease precision medicine with ethical and regulatory perspectives

CRISPR-Based Genome Editing in Contemporary Clinical Medicine: Therapeutic Translation, Global Challenges and the Future of Precision Medicine

CRISPR has progressed from an experimental genome-engineering technology to a clinically relevant therapeutic platform, although its future impact will depend on the ability to combine molecular precision and durable therapeutic benefit with rigorous safety assessment, responsible governance, and equitable access across diverse populations and healthcare systems.

FROM ONE-SIZE-FITS-ALL TO ON-DEMAND: PERSONALIZED CRISPR GENE EDITING FOR RARE GENETIC LIVER DISEASES.

This review provides a comprehensive and critical appraisal of the current landscape of CRISPR-based therapies for genetic liver diseases, and underscores that CRISPR gene editing is transitioning from experimental promise to clinical reality for genetic liver diseases, with personalized approaches poised to redefine the treatment paradigm.

Recent Advances in CRISPR/Cas Technologies for Biological Discovery, Therapeutics, and Diagnostics

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system began as a tool for programmable genome editing. CRISPR/ Cas technologies have evolved into a versatile platform for functional genomic screening, epigenome editing, therapeutic target discovery, and highly sensitive molecular diagnostics. New effectors and engineered variants continue to push these applications into personalized medicine and point-of-care testing. Here, this review highlights recent advances in therapeutic target discovery and therapeutic and molecular diagnostic development utilizing CRISPR/Cas technologies. We discuss how CRISPR interference (CRISPRi), CRISPR activation (CRISPRa), base editing, and prime editing have improved our understanding of disease mechanisms, while creating new opportunities for therapeutic intervention. Current applications in cancer immunotherapy, infectious disease, and neurological disorders are also discussed. In diagnostics, CRISPR-based platforms enable sensitive detection of infectious pathogens, cancer biomarkers, and genetic disorders using programmable nuclease activity in both laboratory and point-of-care settings. Collectively, these advances continue to expand the role of CRISPR/Cas technology across biological discovery, disease diagnostics, and therapeutic development, driving progress in precision medicine.

Toward the clinical application of long-read sequencing in repeat-expansion disorders.

The future of pediatric gene therapy: CRISPR-Cas9, AI, and personalized medicine.

Pediatric gene therapy holds transformative potential to provide curative solutions for children, and the establishment of standardized regulatory frameworks, accompanied by robust and transparent AI integration, will be essential to ensure safe, equitable, and scalable translation.

Towards routine genetic testing of repeat expansions in neurogenetic diseases using multiplex CRISPR-Cas9-targeted long read sequencing

Abnormal expansion of nucleotide repeats was first identified 34 years ago as a unique mutational mechanism. It is now linked to numerous neurogenetic disorders, several of which discovered only recently. The identification of these expansions has led to various classifications based on clinical presentation, repeat nature and genomic location (coding or non-coding regions). Precise diagnosis of these conditions relies on molecular testing, currently performed on a gene-by-gene basis. Their analysis remains challenging, especially for long expansions. We evaluated CRISPR-Cas9-mediated target enrichment coupled to Oxford Nanopore Technologies (ONT) long read sequencing, to accelerate and improve the time-consuming molecular diagnosis of repeat expansion disorders. We simultaneously targeted nine loci involved in 10 repeat expansion disorders in a single capture panel, including FMR1 , HTT , DMPK , CNBP/ZNF9 , ATXN2 , JPH3 , FXN , C9ORF72 and RFC1 , covering a broad range of repeat types, sizes and diagnostic needs. Results were compared with standard routine testing methods. ONT sequencing using Flongle flow cells yielded results consistent with standard techniques for most loci, particularly for non-complex repeats. However, limitations were observed for structurally complex regions such as RFC1 , and inter-run variability required the aggregation of multiple Flongle runs per sample to achieve robust genotyping. These findings highlight both the potential and current limitations of CRISPR-Cas9-enriched ONT sequencing for multiplex diagnosis of repeat expansion disorders in a clinical setting. The approach deserves further development, particularly optimisation of protocols, inclusion of larger sample sizes, and comparison with alternative technologies.