Small molecules for increasing precise genome editing efficiency
Inventors
Riesenberg, Stephan • Maricic, Tomislav
Assignees
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Publication Number
US-12686875-B2
Publication Date
2026-07-21
Expiration Date
Abstract
The present invention relates to compounds suitable to increase precise genome editing efficiency in a eukaryotic target cell or target organism. Thus, the present invention can be applied in gene therapy.
Core Innovation
The patent describes small-molecule compounds, including Nedisertib (M3814), that act as DNA-PKcs inhibitors to increase homologous recombination (HDR)-mediated genome-editing efficiency. The genome editing is performed in eukaryotic genome editing target cells and eukaryotic genome editing target organisms that comprise a DNA cleavage enzyme selected from CRISPR/Cas9, a mutated nickase version of CRISPR/Cas9, a CRISPR/Cpf1 enzyme, or a split-fusion version of any of the foregoing.
The patent frames the problem as improving the outcome of genome editing by shifting the balance among repair pathways. It contrasts HDR with other pathways including non-homologous end joining (NHEJ) and alternative end-joining routes such as microhomology-mediated end joining (MMEJ) and single strand annealing (SSA), and it discusses prior approaches using known DNA-PKcs inhibitors and prior small-molecule enhancers including SCR7, NU7026, NU7441, RS-1, and L755507.
Beyond DNA-PKcs inhibition, the patent includes combinations that modulate additional repair pathways to affect outcomes such as indels and the proportion of desired editing. It describes combining Nedisertib (M3814) with inhibition of alternative end-joining pathways, including inhibition of POLQ/PoIQ for MMEJ and inhibition of RAD52 for SSA, to reduce indels and strongly increase HDR for targets prone to MMEJ or long-homology-driven indel formation.
Claims Coverage
The partial content provided includes two independent method claims. The main inventive features are concentrated in introducing Nedisertib (M3814) as a DNA-PKcs inhibitor to improve HDR-mediated genome editing efficiency under equivalent conditions.
Introducing Nedisertib (M3814) to increase HDR-mediated genome-editing efficiency versus known DNA-PKcs inhibitors
A method comprising introducing Nedisertib (M3814) or a physiologically acceptable salt or solvate into a eukaryotic genome editing target cell or eukaryotic genome editing target organism, wherein the target comprises a DNA cleavage enzyme selected from CRISPR/Cas9, a mutated nickase version of CRISPR/Cas9, a CRISPR/Cpf1 enzyme, or a split-fusion version of any of the foregoing, wherein introducing Nedisertib (M3814) or the physiologically acceptable salt or solvate results in an increase in HDR-mediated genome-editing efficiency greater than that achieved by treatment with known DNA-PKcs inhibitors under equivalent conditions.
Introducing Nedisertib (M3814) to increase HDR-mediated genome-editing efficiency in vitro versus known DNA-PKcs inhibitors
A method for editing a genome of a eukaryotic genome editing target cell in vitro comprising introducing Nedisertib (M3814) or a physiologically acceptable salt or solvate into the eukaryotic genome editing target cell, wherein the cell comprises a DNA cleavage enzyme selected from CRISPR/Cas9, a mutated nickase version of CRISPR/Cas9, a CRISPR/Cpf1 enzyme, or a split-fusion version of any of the foregoing, wherein introducing Nedisertib (M3814) or the physiologically acceptable salt or solvate produces a HDR-mediated genome-editing efficiency greater than that obtained with known DNA-PKcs inhibitors under equivalent conditions.
Across the independent claims, coverage centers on introducing Nedisertib (M3814) or a physiologically acceptable salt or solvate into genome editing target eukaryotic cells or organisms containing specified CRISPR/Cas9 forms and/or CRISPR/Cpf1 enzymes, where HDR-mediated genome-editing efficiency is increased beyond that achieved by known DNA-PKcs inhibitors under equivalent conditions. The in vitro claim narrows the method to eukaryotic genome editing target cells.
Stated Advantages
Increases HDR-mediated genome-editing efficiency compared with treatment with known DNA-PKcs inhibitors under equivalent conditions.
Produces HDR-mediated genome-editing efficiency greater than that obtained with known DNA-PKcs inhibitors under equivalent conditions.
Documented Applications
In vitro genome editing of a eukaryotic genome editing target cell using specified CRISPR/Cas9 variants or CRISPR/Cpf1, with Nedisertib (M3814) introduced to increase HDR-mediated genome-editing efficiency.
Genome editing of a eukaryotic genome editing target cell or a eukaryotic genome editing target organism using specified CRISPR/Cas9 variants or CRISPR/Cpf1, with Nedisertib (M3814) introduced to increase HDR-mediated genome-editing efficiency.
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