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A systematic review and meta-analysis of CRISPR-Cas9 off-target effects in models of blood-related genetic diseases
University of Skövde, School of Bioscience.
2026 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The CRISPR-Cas9 system represents a transformative therapeutic platform for monogenic blood disorders such as sickle cell disease (SCD) and β-thalassemia. However, clinical translation is limited by safety concerns related to unintended off-target genomic alterations. This systematic review and meta-analysis quantified the prevalence and evaluated the severity of CRISPR-Cas9 off-target effects in hematopoietic models of blood-related genetic diseases. Seventy-eight studies were included in qualitative synthesis, and five comparative studies were analysed using a random-effects meta-analysis. High-fidelity Cas9 variants significantly reduced off-target risk compared with wild-type SpCas9 (pooled OR = 0.15, 95% CI: 0.08–0.29), corresponding to an 85% reduction in off-target odds. Substantial heterogeneity was observed (I² = 76.3%), primarily attributable to differences in off-target detection methods, as variations in methodological sensitivity can systematically influence the number and frequency of detected off-target events, contributing substantially to the observed heterogeneity. Subgroup analyses showed greater specificity improvements when assessed using unbiased genome-wide methods (OR = 0.08) compared with targeted sequencing (OR = 0.23). Although high-fidelity variants markedly improve safety profiles, methodological variability and limited long-term in vivo data remain important considerations. These findings provide quantitative evidence supporting the potential use of high-fidelity nucleases and comprehensive genome-wide assessment to enhance the safety of CRISPR-based therapies for hemoglobinopathies.

Place, publisher, year, edition, pages
2026. , p. 34
National Category
Medical Bioscience
Identifiers
URN: urn:nbn:se:his:diva-26373OAI: oai:DiVA.org:his-26373DiVA, id: diva2:2061341
Subject / course
Bioscience
Educational program
Molecular Biotechnology - Master’s Programme, 60 ECTS
Supervisors
Examiners
Available from: 2026-05-20 Created: 2026-05-20 Last updated: 2026-05-20Bibliographically approved

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CiteExportLink to record
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