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AssigneePioneer Hi-Bred International, Inc.
InventorsZHENGLIN HOU, JOSHUA K YOUNG, GIEDRIUS GASIUNAS, VIRGINIJUS SIKSNYS
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Compositions and methods are provided for novel Cas9 orthologs, including, but not limiting to, novel guide polynucleotide/Cas9 endonucleases complexes, single or dual guide RNAs, guide RNA elements, and Cas9 endonucleases. The present disclosure also describes methods for creating a double strand break in a target polynucleotide, methods for genome modification of a target sequence under various in vivo and in vitro conditions, in the genome of a cell, for gene editing, and for inserting a polynucleotide of interest into the genome of a cell. Also provided are nucleic acid constructs and cells having a modified target site or altered polynucleotide of interest produced by the methods described herein.
A synthetic composition comprising a heterologous component and a Cas endonuclease, wherein the Cas endonuclease comprises at least one amino acid feature selected from the group consisting of:
A Cas endonuclease or deactivated Cas endonuclease that recognizes a PAM selected from the group consisting of:
A method of detecting a target polynucleotide sequence, comprising:
A method of binding a Cas endonuclease and guide polynucleotide complex to a target polynucleotide, comprising:
A method of creating a double strand break in a target polynucleotide, comprising:
A method for editing the genome of a cell, the method comprising providing to the cell:
A method of modulating the expression of a gene in a cell, the method comprising providing to the cell:
A method of editing at least one base of a target polynucleotide, comprising:
A method of editing a plurality of bases of a target polynucleotide, comprising:
A method of optimizing the activity of a Cas molecule comprising introducing at least one nucleotide modification to a sequence that comprises at least one amino acid feature selected from the group consisting of:
A method of optimizing the activity of a Cas9 molecule by subjecting a parental Cas9 molecule to at least one round of stochastic protein shuffling, and selecting a resultant molecule that has at least one characteristic not present in the parental Cas9 molecule;
A method of optimizing the activity of a Cas9 molecule by subjecting a parental Cas9 molecule to at least one round of non-stochastic protein shuffling, and selecting a resultant molecule that has at least one characteristic not present in the parental Cas9 molecule;
Dependencies are inferred from the claim text.