The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Apr. 08, 2025

Filed:

Aug. 05, 2022
Applicant:

European Molecular Biology Laboratory, Heidelberg, DE;

Inventors:

Orsolya Barabas, Gaiberg, DE;

Irma Querques, Zurich, CH;

Cecilia Ines Zuliani, Heidelberg, DE;

Assignee:
Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12N 9/12 (2006.01);
U.S. Cl.
CPC ...
C12N 9/1241 (2013.01);
Abstract

This disclosure provides improved reagents and methods for inserting a transgene into the genome of a living host cell. Sleeping beauty (SB) transposase is used in protein form rather than as a vector. This has been made possible by using rational mutagenesis in a particular region of the SB crystal structure to develop variant SB transposase protein that is more soluble. The changes increase solubility while maintaining transposase activity, thereby adapting the variant transposase for the purpose of promoting recombinant integration of a transgene into a target cell when used in protein form rather than as a polynucleotide vector. The modified transposase is highly soluble in electroporation buffer and thermostable during storage. When introduced into a host cell, it promotes integration of a transgene into the genome of the cell in a dose-dependent manner. It is degraded within 48 hours, thereby rapidly clearing transposase activity from the host cell.


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