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US 11,959,073 B2Granted
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.
Claims20
A method of increasing mutagenesis at a double-strand DNA (dsDNA) break at a selected dsDNA target site in a eukaryotic cell comprising:
- a) selecting a dsDNA target site for mutagenesis;
- and
- b) introducing into the eukaryotic cell a polynucleotide sequence encoding a homing endonuclease that binds and cleaves the selected dsDNA target site, and an exonuclease;
- wherein the exonuclease exhibits exonuclease activity at the cleaved dsDNA target site, resulting in increased mutagenesis at the selected dsDNA target site as compared to mutagenesis that occurs in the absence of exonuclease activity.
1 more element
Dependencies are inferred from the claim text.