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Journal Article

Efficient high-precision homology-directed repair-dependent genome editing by HDRobust

MPS-Authors
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Riesenberg,  Stephan       
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;
The Leipzig School of Human Origins (IMPRS), Max Planck Institute for Evolutionary Anthropology, Max Planck Society;
Genome Engineering and Repair, Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Kanis,  Philipp       
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;
The Leipzig School of Human Origins (IMPRS), Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Macak,  Dominik
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Wollny,  Damian       
Single Cell Genomics, Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Düsterhöft,  Dorothee
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Kowalewski,  Johannes
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Helmbrecht,  Nelly       
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;
The Leipzig School of Human Origins (IMPRS), Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Maricic,  Tomislav       
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Pääbo,  Svante       
Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;
The Leipzig School of Human Origins (IMPRS), Max Planck Institute for Evolutionary Anthropology, Max Planck Society;
Neandertals and more, Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Max Planck Society;

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Riesenberg_Efficient_NatMeth_2023.pdf
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Citation

Riesenberg, S., Kanis, P., Macak, D., Wollny, D., Düsterhöft, D., Kowalewski, J., et al. (2023). Efficient high-precision homology-directed repair-dependent genome editing by HDRobust. Nature Methods, 20, 1388-1399. doi:10.1038/s41592-023-01949-1.


Cite as: https://hdl.handle.net/21.11116/0000-000D-78D4-D
Abstract
Homology-directed repair (HDR), a method for repair of DNA
double-stranded breaks can be leveraged for the precise introduction
of mutations supplied by synthetic DNA donors, but remains limited by
low efficiency and off-target effects. In this study, we report HDRobust,
a high-precision method that, via the combined transient inhibition of
nonhomologous end joining and microhomology-mediated end joining,
resulted in the induction of point mutations by HDR in up to 93% (median
60%, s.e.m. 3) of chromosomes in populations of cells. We found that, using
this method, insertions, deletions and rearrangements at the target site, as
well as unintended changes at other genomic sites, were largely abolished.
We validated this approach for 58 different target sites and showed that it
allows efficient correction of pathogenic mutations in cells derived from
patients suffering from anemia, sickle cell disease and thrombophilia.