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US 2023/0416711 A1Published application
Rationally-designed single-chain meganucleases with non-palindromic recognition sequences
AssigneePrecision BioSciences, Inc.
InventorsJames Jefferson Smith, Derek Jantz
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Abstract
Disclosed are rationally-designed, non-naturally-occurring meganucleases in which a pair of enzyme subunits having specificity for different recognition sequence half-sites are joined into a single polypeptide to form a functional heterodimer with a non-palindromic recognition sequence. The invention also relates to methods of producing such meganucleases, and methods of producing recombinant nucleic acids and organisms using such meganucleases.
Claims18
-37. (canceled)
A recombinant single-chain meganuclease comprising:
- a first LAGLIDADG subunit derived from a first mono-LAGLIDADG meganuclease, said first LAGLIDADG subunit having a first recognition half-site;
- a second LAGLIDADG subunit derived from a second mono-LAGLIDADG meganuclease or a di-LAGLIDADG meganuclease, said second LAGLIDADG subunit having a second recognition half-site;
- wherein said first and second LAGLIDADG subunits are covalently joined by a polypeptide linker such that said first LAGLIDADG domain is N-terminal to said linker and said second LAGLIDADG domain is C-terminal to said linker;
- and
- wherein said first and second LAGLIDADG subunits are capable of functioning together to recognize and cleave a non-palindromic DNA sequence which is a hybrid of said first recognition half-site and said second recognition half-site.
2 more elements
Dependencies are inferred from the claim text.