The Role of Miniature Cas12 Orthologs in In Vivo Gene Editing: A Systematic Review of Current Applications and Future Prospects

Introduction: Miniature Cas12 orthologs have recently gained attention for their compact size and efficiency, making them ideal for in vivo gene-editing applications, especially with adeno-associated viral (AAV) vectors. This systematic review explores the current uses of miniature Cas12 orthologs in gene-editing studies, highlighting their benefits and potential challenges for future clinical applications.

Methods: A comprehensive literature search was conducted to identify relevant studies published from 2015 to 2023. Studies focusing on the use of miniature Cas12 orthologs for in vivo gene editing were selected. Data extraction included parameters such as gene-editing efficiency, delivery methods, off-target effects, and therapeutic outcomes. Statistical analyses using Python and R were performed to compare the effectiveness of various Cas12 orthologs across different organisms and disease models, including a meta-analysis of editing success rates.

Results: A total of 35 studies involving miniature Cas12 orthologs were included. These orthologs showed an average gene-editing efficiency of 78% across all models, with AAV vectors being the most commonly used delivery method (67%). Off-target effects were observed in only 3% of cases, indicating high specificity compared to larger Cas variants. In therapeutic applications, Cas12 orthologs were particularly effective in liver and muscular dystrophy models, achieving significant phenotypic improvement in 82% of cases. Meta-analysis confirmed the higher efficiency of miniature Cas12 orthologs in models with compact gene targets, with no significant loss of editing performance.

Conclusion : Miniature Cas12 orthologs represent a promising advancement in in vivo gene editing, combining efficiency and specificity with AAV vector compatibility. These findings support their continued development for clinical applications, especially in diseases requiring targeted and precise gene correction. Future research should focus on expanding their applicability to larger and more complex genomes, as well as further improving their safety profiles.