Improving DNA repair efficiency for CRISPR/Cas9-mediated genome editing
Inventors: Bruna Paulsen, Pankaj K Mandal, Paula Guitierrze-Martinez, Derrick Rossi, Wataru Ebina
Invention Types: Therapeutics
Research Areas: Genetic Disease, Personalized Medicine, Stem Cell
Keywords: Gene, Gene TherapyFor More Information Contact: Ives, Catherine L.
Gene editing approaches have the potential to transform medicine by treating the root of the problem (i.e., gene mutation) in genetic diseases rather than the symptoms. However, a major challenge in the path towards clinical translation is the ability to efficiently repair clinically relevant gene mutations and prevent off-target mutagenesis. CRISPR/Cas9 system is a genome-editing tool for site-directed genome editing in mammalian cells. Gene editing by CRISPR/Cas9 utilizes double-strand brakes generated by Cas9, followed by non-homologous end joining repair. However this may result in small-unintended insertions and deletions, disrupting coding of the targeted gene.
Dr. Rossi and his team found that is possible to improve the gene repair efficiency of CRISPR/Cas9 system through transient expression of Rad 52 and a dominant negative isoform of 53BP1 involved in DNA repair pathways. In studies, his lab found a 3-5-fold increase in repair efficiency at targeted loci at multiple loci in primary cells as well as tissue-specific adult stem cells. This new technique improves the potential of CRISPR/Cas9-mediated genome editing to precisely repair disease-specific somatic mutations and improve its performance in clinical trials.
- Human gene therapy
- Gene-editing tools
- Increase precision in genome editing without altering CRISPR/Cas9 specificity or genome stability
- Technique has the ability to prevent off-target effects
• Mandal et al. Efficient ablation of genes in human hematopoietic stem and effector cells using CRISPR/Cas9. Cell Stem Cell. 2014;15(5):643-52. doi: 10.1016/j.stem.2014.10.004. Epub 2014 Nov 6.
• Paulsen et al. Ectopic expression of RAD52 and dn53BP1 improves homology-directed repair during CRISPR–Cas9 genome editing. Nat. Biomed. Eng. (2017) doi: 10.1038/s41551-017-0145-2.
IPStatus: Pat. Pend.