Activation of homology-directed DNA repair plays key role in CRISPR-mediated genome correction.

Autor: Mondal G; Department of Surgery, Mayo Clinic, Rochester, MN, USA., VanLith CJ; Department of Surgery, Mayo Clinic, Rochester, MN, USA., Nicolas CT; Department of Surgery, Mayo Clinic, Rochester, MN, USA.; Department of Surgery, University of Alabama Birmingham, Birmingham, AL, USA., Thompson WS; Department of Pediatric and Adolescent Medicine, Mayo Clinic, Rochester, MN, USA., Cao WS; Department of Surgery, Mayo Clinic, Rochester, MN, USA., Hillin L; Department of Surgery, Mayo Clinic, Rochester, MN, USA., Haugo BJ; Department of Surgery, Mayo Clinic, Rochester, MN, USA., Brien DRO; Department of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN, USA., Kocher JP; Department of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN, USA., Kaiser RA; Department of Surgery, Mayo Clinic, Rochester, MN, USA.; Midwest Fetal Care Center, Children's Hospital of Minnesota, Minneapolis, MN, USA., Lillegard JB; Department of Surgery, Mayo Clinic, Rochester, MN, USA. jlillegard@msn.com.; Midwest Fetal Care Center, Children's Hospital of Minnesota, Minneapolis, MN, USA. jlillegard@msn.com.; Pediatric Surgical Associates, Minneapolis, MN, USA. jlillegard@msn.com.
Jazyk: angličtina
Zdroj: Gene therapy [Gene Ther] 2023 Apr; Vol. 30 (3-4), pp. 386-397. Date of Electronic Publication: 2022 Oct 19.
DOI: 10.1038/s41434-022-00369-8
Abstrakt: Gene editing for the cure of inborn errors of metabolism (IEMs) has been limited by inefficiency of adult hepatocyte targeting. Here, we demonstrate that in utero CRISPR/Cas9-mediated gene editing in a mouse model of hereditary tyrosinemia type 1 provides stable cure of the disease. Following this, we performed an extensive gene expression analysis to explore the inherent characteristics of fetal/neonatal hepatocytes that make them more susceptible to efficient gene editing than adult hepatocytes. We showed that fetal and neonatal livers are comprised of proliferative hepatocytes with abundant expression of genes involved in homology-directed repair (HDR) of DNA double-strand breaks (DSBs), key for efficient gene editing by CRISPR/Cas9. We demonstrated the same is true of hepatocytes after undergoing a regenerative stimulus (partial hepatectomy), where post-hepatectomy cells show a higher efficiency of HDR and correction. Specifically, we demonstrated that HDR-related genome correction is most effective in the replicative phase, or S-phase, of an actively proliferating cell. In conclusion, this study shows that taking advantage of or triggering cell proliferation, specifically DNA replication in S-phase, may serve as an important tool to improve efficiency of CRISPR/Cas9-mediated genome editing in the liver and provide a curative therapy for IEMs in both children and adults.
(© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)
Databáze: MEDLINE
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