# US 10,767,169 B1 · Nucleic acid-guided nucleases

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- Number: US 10,767,169 B1
- Document ID: US10767169B1
- Status: Granted
- Kind: B1
- Application number: 16/928,742
- Filed: 2020-07-14
- Granted: 2020-09-08
- Earliest claimed: 2019-12-13
- Assignees: Inscripta, Inc.
- Applicants: Inscripta, Inc.
- Inventors: Juhan Kim; Benjamin Mijts; Aamir Mir; Andrew Garst; Kyle Seamon
- Web page: https://patentgrep.com/patent/US10767169B1

## CPC

- C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]}
- C12N 9/222: {Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes}
- C12N 15/102: {Mutagenizing nucleic acids}
- C12N 15/70: Vectors or expression systems specially adapted for E. coli
- C12N 15/74: Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N 15/81: for yeasts
- C12N 15/85: for animal cells
- C12N 15/905: {in yeast}
- C12N 15/907: {in mammalian cells}
- C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR]
- C12N 2310/531: Stem-loop; Hairpin
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure provides novel RNA-guided enzymes for making rational and direct edits to the genome of live cells.

## Claims

### Claim 1

A coding sequence for a nucleic acid-guided nuclease comprising a nucleic acid sequence of any of SEQ ID Nos. 20, 21, 22 or 24.

### Claim 2 (depends on 1)

The coding sequence for the nucleic acid-guided nuclease of claim 1 comprising the nucleic acid sequence SEQ ID No. 20.

### Claim 3 (depends on 2)

The coding sequence for the nucleic acid-guided nuclease of claim 2, wherein a compatible guide RNA to the nucleic-acid-guided nuclease has an optimal crRNA variable loop comprising UCUU.

### Claim 4 (depends on 1)

The coding sequence for the nucleic acid-guided nuclease of claim 1 comprising the nucleic acid sequence SEQ ID No. 21.

### Claim 5 (depends on 4)

The coding sequence for the nucleic acid-guided nuclease of claim 4, wherein a compatible guide RNA to the nucleic-acid-guided nuclease has an optimal crRNA variable loop comprising UCUU or UGUU.

### Claim 6 (depends on 1)

The coding sequence for the nucleic acid-guided nuclease of claim 1 comprising the nucleic acid sequence SEQ ID No. 22.

### Claim 7 (depends on 6)

The coding sequence for the nucleic acid-guided nuclease of claim 6, wherein a compatible guide RNA to the nucleic-acid-guided nuclease has an optimal crRNA variable loop comprising UCUU or UAUU.

### Claim 8 (depends on 1)

The coding sequence for the nucleic acid-guided nuclease of claim 1 comprising the nucleic acid sequence SEQ ID No. 24.

### Claim 9 (depends on 8)

The coding sequence for the nucleic acid-guided nuclease of claim 8, wherein a compatible guide RNA to the nucleic-acid-guided nuclease has an optimal crRNA variable loop comprising UAUU.

### Claim 10 (depends on 1)

The coding sequence for a nucleic acid-guided nuclease of claim 1 for editing in bacteria.

### Claim 11 (depends on 1)

The coding sequence for a nucleic acid-guided nuclease of claim 1 for editing in yeast.

### Claim 12 (depends on 1)

The coding sequence for a nucleic acid-guided nuclease of claim 1 for editing in isolated mammalian cells.