# US 11,441,132 B2 · Mutated sleeping beauty transposase

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- Number: US 11,441,132 B2
- Document ID: US11441132B2
- Status: Granted
- Kind: B2
- Application number: 16/640,976
- Filed: 2018-08-17
- Published: 2021-04-29
- Granted: 2022-09-13
- Earliest claimed: 2017-08-21
- Assignees: European Molecular Biology Laboratory
- Applicants: European Molecular Biology Laboratory
- Inventors: Orsolya Barabas; Irma Querques; Cecilia Ines Zuliani
- Web page: https://patentgrep.com/patent/US11441132B2

## CPC

- C12N 9/1241: {Nucleotidyltransferases (2.7.7)}
- 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 invention relates to improved transposase polypeptides having increased solubility. The enzyme of the invention was developed based on the Sleeping Beauty (SB) transposase. The invention provides further nucleic acids, vectors and recombinant cells encoding or containing the improved transposase, as well as a transposase system. Furthermore provided are medical and non-medical uses of the transposase of the invention for gene delivery. The invention is in particular useful as a tool for gene delivery in genetically modified cell based therapeutic approaches for treating various diseases.

## Claims

### Claim 1

A transposase polypeptide, wherein the transposase polypeptide is a Sleeping Beauty (SB) transposase and comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:2 but with at least one mutated amino acid residue at position 176 or position 212 of SEQ ID NO:2.

### Claim 2 (depends on 1)

The transposase polypeptide according to claim 1, wherein the transposase polypeptide comprises a mutation at position 176 and a mutation at position 212 of SEQ ID NO:2.

### Claim 3 (depends on 1)

The transposase polypeptide according to claim 1, wherein the at least one mutated amino acid residue is mutated into a serine residue.

### Claim 4 (depends on 1)

The transposase polypeptide according to claim 1, wherein the amino acid sequence has at least 90% sequence identity to an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3 between residues 150 to 250.

### Claim 5 (depends on 1)

A transposon system comprising (a) a transposon unit containing inverted terminal repeats (ITRs) or direct terminal repeats (DTRs) that flank a sequence of interest to be inserted into the genome of a target cell; and (b) a transposase polypeptide according to claim 1.

### Claim 6 (depends on 1)

A pharmaceutical composition comprising a transposase polypeptide according to claim 1, together with a pharmaceutically acceptable carrier and/or excipient.

### Claim 7 (depends on 1)

A kit comprising (a) a transposon unit containing inverted terminal repeats (ITRs) or DTRs that flank a sequence of interest to be inserted into the genome of a target cell; and (b) a transposase polypeptide according to claim 1.

### Claim 8 (depends on 2)

The transposase polypeptide according to claim 2, wherein the transposase polypeptide comprises mutations C176S and I212S.

### Claim 9 (depends on 4)

The transposase polypeptide according to claim 4, wherein the amino acid sequence has at least 90% sequence identity to the full-length sequence shown in SEQ ID NO: 1 or SEQ ID NO: 3.