Methods and compositions for polymerase ii (pol-ii) based guide rna expression
AssigneeDanisco US Inc.
InventorsFelipe Oseas Bendezu, Xiaochun Fan, Ryan L. Frisch, Seung-Pyo Hong
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Abstract
Compositions and methods are provided for editing nucleotides and/or altering target sites in the genome of a cell. The methods and compositions employ a recombinant DNA construct comprising a Pol-II promoter operably linked to a polynucleotide encoding a single guide RNA, wherein said guide RNA is capable of forming a guide RNA/Cas endonuclease complex, wherein said complex can bind to and cleave a target site sequence in the genome of a cell such as a eukaryote cell. The present disclosure further describes methods and compositions employing said recombinant DNA construct to modify the genome of non-conventional yeast.
Claims16
A recombinant DNA construct comprising a Polymerase II (Pol-II) promoter operably linked to a polynucleotide encoding a single guide RNA, wherein said guide RNA is capable of forming a guide RNA/Cas endonuclease complex, wherein said complex can bind to and cleave a target site sequence in the genome of a eukaryote.
A recombinant DNA construct comprising a Polymerase II (Pol-II) promoter operably linked to a polynucleotide encoding a dual guide RNA (crRNA and tracrRNA), wherein the dual guide RNA is capable of forming a guide RNA/Cas endonuclease complex, wherein said complex can bind to and cleave a target site sequence in the genome of a eukaryote.
Dependencies are inferred from the claim text.