Genome-Editing-Technologies - Single compound or compound mix dramatically improve Homology-Directed-Repair (HDR)

Scientists at the Max-Planck Institute for Evolutionary Anthropology in Leipzig have developed
two approaches to drastically enhance HDR efficiency by blocking NHEJ. Both do block kinases (DNA-PKcs) chemically or genetically (DNA-PKc mutant) that are promoting NHEJ.

1. iCRISPY compound mix. Such mixture contains several inhibitors of DNA-PKcs (our reference: MI-5288) - or even just one (“M-compound-only” still enabling strong HDR; our reference: MI-5734). Both approaches enable strong and recently unseen HDR efficiencies up to 90% in cell culture. iCRISPY mix significantly increases HDR rates independently of cell types or CRISPR enzymes used, when transiently applied.
2. Genetically modified cell lines harbouring mutated DNA-PKcs permit high HDR rates while confidently maintaining genome stability. DNA-PKc mutant cell lines from HEK293 cells, K562 cells and human pluripotent stem cells (409B2) are readily available.

• MI 1306-5288-LI: see WO18/189186 ;

• MI 1306-5734-LI: see WO20/127738

1. Targeting repair pathways with small molecules increases precise genome editing in pluripotent stem cells, Riesenberg and Maricic, Nat Commun. 2018 Jun 4;9(1):2164
2. Simultaneous precise editing of multiple genes in human cells, Nucleic Acids Res. Riesenberg et al., 2019 Nov 4;47(19):e116