Mutations that Enhance the DNA Cleavage Activity of Acidaminococcus Sp. CPF1
AssigneeIntegrated Dna Technologies, Inc.
InventorsLiyang ZHANG, Christopher Anthony VAKULSKAS, Nicole Mary BODE, Michael Allen COLLINGWOOD, Kristin Renee BELTZ, Mark Aaron BEHLKE
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Abstract
The present disclosure concerns polynucleotides and amino acids of Acidaminococcus sp. Cas12a (Cpf1) and methods for their use for genome editing in eukaryotic cells.
Claims21
1-41. (canceled)
A method of performing therapeutic gene editing in a eukaryotic cell, comprising, introducing a CRISPR/Cas endonuclease system into the eukaryotic cell, wherein the CRISPR/Cas endonuclease system comprises:
- i. a nucleic acid encoding a Cas12a variant, wherein the Cas12a variant comprises one or more substitutions selected from the group consisting of M537R, F870L, and M537R/F870L substitutions relative to the wild-type Cas12a sequence of SEQ ID NO:
- 462;
- and
- ii. at least one guide RNA that hybridizes to a target sequence of a DNA molecule in a eukaryotic cell,
- wherein the eukaryotic cell is selected from the group consisting of human cells, and non-human animal cells, further wherein the target sequence of the DNA molecule in the eukaryotic cell is edited.
An mRNA encoding a Cas12a variant, wherein the Cas12a variant comprises one or more substitutions selected from the group consisting of M537R, F870L, and M537R/F870L substitutions relative to the wild-type Cas12a sequence of SEQ ID NO:
- 462.
Dependencies are inferred from the claim text.