# US 11,306,298 B1 · Mad nucleases

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- Number: US 11,306,298 B1
- Document ID: US11306298B1
- Status: Granted
- Kind: B1
- Application number: 17/463,498
- Filed: 2021-08-31
- Granted: 2022-04-19
- Earliest claimed: 2021-01-04
- Assignees: Inscripta, Inc.
- Applicants: Inscripta, Inc.
- Inventors: Juhan Kim; Benjamin Mijts; Aamir Mir
- Web page: https://patentgrep.com/patent/US11306298B1

## CPC

- C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]}
- C12N 9/222: {Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes}
- C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity}
- C12N 15/113: 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)}
- C12N 15/907: {in mammalian cells}
- C12Y 301/21004: Type II site-specific deoxyribonuclease (3.1.21.4)
- C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR]
- C12N 2320/50: Methods for regulating/modulating their activity
- C12N 2800/80: Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The present disclosure provides new RNA-guided nuclease systems and engineered nickases for making rational, direct edits to nucleic acids in live cells.

## Claims

### Claim 1

A nickase selected from the following nickases: MAD2016-H851A [SEQ ID NO: 177]; MAD2016-N874A [SEQ ID NO: 178]; MAD2032-H590A [SEQ ID NO: 179]; MAD-2039-H587A [SEQ ID NO: 180]; MAD2039-N610A [SEQ ID NO: 181].

### Claim 2 (depends on 1)

The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 177.

### Claim 3 (depends on 1)

The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 178.

### Claim 4 (depends on 1)

The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 179.

### Claim 5 (depends on 1)

The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 180.

### Claim 6 (depends on 1)

The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 181.