Compositions And Methods Of Nucleic Acid-Targeting Nucleic Acids

Patent metadata

Field	Value
Publication number	US-2016251640-A1
Application number	US-201615159776-A
Country	US
Kind code	A1
Filing date	May 19, 2016
Priority date	Mar 14, 2013
Publication date	Sep 1, 2016
Grant date	—

Abstract

This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.

First claim

1 - 26 . (canceled)

27 . An engineered CRISPR Cas9 protein comprising:
a modification in a region of a CRISPR Cas9 protein corresponding to a region of amino acid residues 170-312 of a S. pyogenes -Cas9 protein (SEQ ID NO:8), wherein the modification comprises an insertion, deletion, or combination thereof.

28 . The engineered CRISPR Cas9 protein of claim 27 , wherein the modification is made in identified regions of sequence similarity between the Cas9 protein and the S. pyogenes -Cas9 protein (SEQ ID NO:8).

29 . The engineered CRISPR Cas9 protein of claim 27 , wherein the modification comprises a deletion.

30 . The engineered CRISPR Cas9 protein of claim 27 , wherein the modification comprises a deletion of more than 5 amino acid residues.

31 . The engineered CRISPR Cas9 protein of claim 27 , wherein the modification comprises a deletion of more than 100 amino acid residues.

32 . The engineered CRISPR Cas9 protein of claim 27 , wherein the modification comprises a deletion and an insertion of a non-native amino acid sequence.

33 . The engineered CRISPR Cas9 protein of claim 27 , wherein the modification comprises an insertion of a non-native amino acid sequence to form a fusion protein.

34 . The engineered CRISPR Cas9 protein of claim 33 , wherein the non-native amino acid sequence comprises a functional domain.

35 . The engineered CRISPR Cas9 protein of claim 27 , wherein the engineered CRISPR Cas9 protein is a fusion.

36 . The engineered CRISPR Cas9 protein of claim 35 , wherein the engineered CRISPR Cas9 fusion comprises a detectable label.

37 . The engineered CRISPR Cas9 protein of claim 35 , wherein the engineered CRISPR Cas9 fusion comprises the engineered CRISPR Cas9 protein and an endonuclease.

38 . The engineered CRISPR Cas9 protein of claim 37 , wherein the endonuclease is FokI.

39 . The engineered CRISPR Cas9 protein of claim 27 , wherein the engineered CRISPR Cas9 protein is enzymatically inactive.

40 . The engineered CRISPR Cas9 protein of claim 27 , wherein the engineered CRISPR Cas9 protein is codon optimized for a host cell of interest.

41 . The engineered CRISPR Cas9 protein of claim 40 , wherein the host cell of interest is a bacterial cell, an archaeal cell, a cell of a single-cell eukaryotic organism, a plant cell, an algal cell, a fungal cell, an animal cell, a cell from an invertebrate animal, a cell from a vertebrate animal, or a cell from a mammal.

42 . The engineered CRISPR Cas9 protein of claim 41 , wherein the host cell is a human cell.

43 . A polynucleotide encoding the engineered CRISPR Cas9 protein of claim 27 .

44 . The polynucleotide of claim 43 , wherein the modification of the engineered CRISPR Cas9 protein is a deletion of more than 5 amino acid residues.

45 . The polynucleotide of claim 43 , wherein the modification of the engineered CRISPR Cas9 protein is a deletion of more than 100 amino acid residues.

46 . A kit comprising:
the engineered CRISPR Cas9 protein of claim 27 or a polynucleotide encoding the engineered CRISPR Cas9 protein; and a reagent.

Assignees

• Caribou Biosciences Inc

Inventors

• May Andrew Paul
• Haurwitz Rachel E
• Doudna Jennifer A
• Berger James M
• Carter Matthew Merrill
• Donohoue Paul Daniel

Classifications

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/04

  Antibacterial agents · CPC title

• A61P27/02

  Ophthalmic agents · CPC title