Novel small molecule combinations that favour CRISPR/Cas-mediated homology-directed repair
CRISPR/Cas9 technology can be used for both gene knock-out and knock-in to reverse the effect of a mutated gene.
DNA double strand breaks (DSBs) are generated by CRISPR/Cas9, which are typically repaired by the competing repair pathways, non-homologous end joining (NHEJ) or homology-directed repair.
Precise genome editing is facilitated by HDR; however, its efficiency is low and the NHEJ pathway is the preferred choice for repair.
Strategies used to increase HDR rates include:
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- Synchronisation of cells (S-phase)
- Overexpression of HDR mediators
- Use of small molecules