Novel crispr-associated systems and components

Patent metadata

Field	Value
Publication number	US-2020299659-A1
Application number	US-202016862261-A
Country	US
Kind code	A1
Filing date	Apr 29, 2020
Priority date	May 16, 2018
Publication date	Sep 24, 2020
Grant date	—

Abstract

The disclosure describes novel systems, methods, and compositions for the manipulation of nucleic acids in a targeted fashion. The disclosure describes non-naturally occurring, engineered Type III-E CRISPR-Cas systems, components, and methods for targeted modification of DNA, RNA, and protein substrates. Each system includes one or more protein components and one or more nucleic acid components that together target DNA, RNA, or protein substrates.

First claim

1 - 55 . (canceled)

56 . An engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)—Cas system comprising:
an RNA guide comprising a direct repeat sequence and a spacer sequence capable of hybridizing to a target nucleic acid; and
a CRISPR-associated protein, wherein the CRISPR-associated protein comprises an amino acid sequence that is at least 95% identical to an amino acid sequence of SEQ ID NO: 316, SEQ ID NO: 317, or SEQ ID NO: 318;
wherein the CRISPR-associated protein is capable of binding to the RNA guide and of targeting the target nucleic acid sequence complementary to the spacer sequence.

57 . The system of claim 56 , wherein the CRISPR-associated protein comprises an amino acid sequence that is at least 95% identical to an amino acid sequence of SEQ ID NO: 316.

58 . The system of claim 56 , wherein the CRISPR-associated protein comprises an amino acid sequence of SEQ ID NO: 316.

59 . The system of claim 56 , wherein the CRISPR-associated protein comprises an amino acid sequence that is at least 95% identical to an amino acid sequence of SEQ ID NO: 317.

60 . The system of claim 56 , wherein the CRISPR-associated protein comprises an amino acid sequence of SEQ ID NO: 317.

61 . The system of claim 56 , wherein the CRISPR-associated protein comprises an amino acid sequence that is at least 95% identical to an amino acid sequence of SEQ ID NO: 318.

62 . The system of claim 56 , wherein the CRISPR-associated protein comprises an amino acid sequence of SEQ ID NO: 318.

63 . The system of claim 56 , wherein the CRISPR-associated protein comprises a RuvC domain.

64 . The system of claim 56 , wherein the CRISPR-associated protein comprises an OrfB_Zn_ribbon domain.

65 . The system of claim 56 , wherein the direct repeat sequence comprises a nucleic acid sequence of TGRGAC (SEQ NO: 319).

66 . The system of claim 56 , wherein the direct repeat sequence comprises a nucleic acid sequence of any of SEQ ID NOS: 321-328, or a sequence having at least 90% identity thereto.

67 . The system of claim 56 , wherein the direct repeat sequence forms a stem-loop.

68 . The system of claim 56 , wherein the spacer sequence has a length of 15-50 nucleotides.

69 . The system of claim 56 , wherein the CRISPR-associated protein is capable of recognizing a protospacer adjacent motif (PAM).

70 . The system of claim 56 , wherein the target nucleic acid is a DNA.

71 . The system of claim 56 , wherein the targeting of the target nucleic acid by the CRISPR-associated protein and RNA guide results in a modification in the target nucleic acid.

72 . The system of claim 56 , wherein the modification in the target nucleic acid is a double stranded cleavage event.

73 . The system of claim 56 , wherein the modification in the target nucleic acid is a single stranded cleavage event.

74 . The system of claim 56 , within a cell.

75 . A method of targeting and editing a target nucleic acid, the method comprising contacting the target nucleic acid with a system of claim 56 .

Assignees

• Arbor Biotechnologies Inc

Inventors

• Cheng David R
• Scott David A
• Yan Winston X

Classifications

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N9/96

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• C12Q1/6832

  Enhancement of hybridisation reaction · CPC title

• C12N2800/80

  Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites · CPC title

• C12N15/907

  in mammalian cells · CPC title