Mutant cpf1 endonucleases
AssigneeUniversity of Copenhagen
InventorsStefano Stella, Guillermo Montoya
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Abstract
The present invention relates to mutant Cpf1 endonucleases having altered activity compared to the wild type Cpf1, and their use to introduce single strand breaks in nucleic acid sequences. Methods for detection and quantification of a nucleic acid sequence are also disclosed. Methods for diagnosis of an infectious disease are also disclosed.
Claims28
1-114. (canceled)
A mutant Cpf1 endonuclease comprising a polypeptide sequence that comprises a deletion or a substitution in a Lid domain sequence, a finger domain sequence, a REC domain sequence, or any combination thereof, wherein the mutant Cpf1 endonuclease exhibits reduced non-specific single-stranded DNase activity compared to wild type Cpf1.
A mutant Cpf1 endonuclease comprising a polypeptide sequence that comprises a deletion or a substitution in a Lid domain sequence, a finger domain sequence, or a REC domain sequence, wherein the mutant Cpf1 endonuclease exhibits non-specific single-stranded DNase activity, and exhibits reduced endonuclease activity on target DNA compared to said endonuclease activity of wild type Cpf1.
A mutant Cpf1 endonuclease comprising a polypeptide sequence that comprises a substitution or deletion of the amino acid residues corresponding to positions 909, 1000, 1001, 1013 and/or 1016 of SEQ ID NO:
- 24 or positions 843, 943, 944, 955 and/or 958 of SEQ ID NO:
- 25, wherein the mutant Cpf1 endonuclease exhibits endonuclease activity upon a target dsDNA sequence to cleave both strands of the target dsDNA, and the mutant Cpf1 endonuclease exhibits reduced non-specific single-stranded DNase activity compared to wild type Cpf1.
A mutant Cpf1 endonuclease comprising a polypeptide sequence that comprises a deletion or a substitution in a Lid domain sequence, a finger domain sequence, or a REC domain sequence, wherein the mutant Cpf1 endonuclease exhibits non-specific single-stranded DNase activity, and exhibits reduced endonuclease activity on target DNA compared to said endonuclease activity of wild type Cpf1.
A mutant Cpf1 endonuclease comprising a polypeptide sequence having at least 95% identity to the sequence of SEQ ID NO:
- 24, wherein the polypeptide comprises at least two amino acid substitutions in the Lid domain sequence corresponding to positions 993-1005 of the amino acid sequence of SEQ ID NO:
- 24, optionally wherein the Lid domain sequence comprises an amino acid substitution at position 1000 and position 1001.
Dependencies are inferred from the claim text.