Coupling endonucleases with end-processing enzymes drives high efficiency gene disruption
AssigneeSeattle Children's Research Institute
InventorsAndrew M. Scharenberg, Michael T. Certo, Kamila Sabina Gwiazda
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Abstract
The present disclosure relates to the co-expression of an endonuclease with an end-processing enzyme for the purpose of enhanced processing of the polynucleotide ends generated by endonuclease cleavage.
Claims36
A fusion polypeptide wherein a homing endonuclease that binds and cleaves a selected dsDNA target site in a mammalian cell is linked by a linker domain to a 3′ to 5′ exonuclease;
- and wherein the homing endonuclease is selected from the group consisting of:
- I-Ltr, I-Gpi, I-Gze, I-MpeMI, I-PanMI, I-CreI, I-OnuI, I-HjeMI, and I-AniI.
A fusion polypeptide wherein a I-CreI homing endonuclease that binds and cleaves a selected dsDNA target site in a mammalian cell is linked by a linker domain to a 3′ to 5′ exonuclease.
A fusion polypeptide wherein a I-OnuI homing endonuclease that binds and cleaves a selected dsDNA target site in a mammalian cell is linked by a linker domain to a 3′ to 5′ exonuclease.
Dependencies are inferred from the claim text.