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Correspondence to Author: Andree-Zeid Kakish*, Bryson Martin*, Meena Adiyodi*, Connor Majewski*, G. Ian Gallicano*. 

Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, 3900 Reservoir Road NW, Washington DC 20007, USA

DOI: 10.52338/joat.2025.5180

Abstract:

For the last ten years, the precision of genome engineering in human embryos has increased tenfold using the CRISPR-Cas9 genome editing tool. Commonly used as an intervention strategy for disease prevention, CRISPR-Cas9’s precision to target disease-carrying genes far exceeds that of its predecessors, ZFNs and TALENs, and has been used for conditions like β-thalassemia, familial hypertrophic cardiomyopathy (FHC), glucose-6-phosphate dehydrogenase (G6PD) deficiency, and many others. As its utilization increases, additions to its procedure to refine its efficacy have also continued, as explained by adding homology-directed repair (HDR), high fidelity Cas9 variants, and chemical enhancers to further increase the success rate, while reducing the off-target effects. Despite current progress, the risk of genomic instability, large deletions, and variable repair mechanisms is still high. Another concern is how the current ethical and regulatory frameworks within the United States do not account for this technological advancement. This review aims to examine current CRISPR-Cas9 methodologies and genetic techniques, to evaluate the associated limitations and challenges, and to explore the ethical implications of this technique in the advancing field.

Keywords: CRISPR-Cas9, Germline Editing, Homology-Directed Repair (HDR), Genomic Instability, Bioethics.

Citation:

Dr. Andree-Zeid Kakish, Genetic Engineering with CRISPR-Cas9: Methodologies, Genetic Techniques, and Bioethics. Journal of Advanced Therapeutics 2025.