公开(公告)号：US20230183713A1

公开(公告)日：2023-06-15

申请号：US18055545

申请日：2022-11-15

Applicant: TECHNISCHE UNIVERSITÄT DRESDEN

Inventor： Jenna HOERSTEN ,  Felix LANSING ,  Frank BUCHHOLZ

IPC: C12N15/52 ,  C12N15/63

CPC classification number: C12N15/52 ,  C12N15/63

Abstract: The invention relates to the field of genome editing and provides a method of generating DNA recombinases or provides DNA recombinases, which efficiently and specifically recombine genomic target sequences as obligate DNA recombinase enzymes. The invention provides a genetically engineered DNA recombining enzyme comprising a complex of a first and a second recombinase enzyme, wherein said first recombinase enzyme and said second recombinase enzyme specifically recognize a first half-site and a second half-site of an upstream target site and/or a downstream target site of a DNA recombinase, wherein said first recombinase enzyme and said second recombinase enzyme each comprises at least one mutation in its catalytic site, wherein said first recombinase enzyme and said second recombinase enzyme carrying said at last one mutation in their catalytic site, when expressed in isolation, do not show the catalytic activity of a DNA recombinase, and wherein said first DNA recombinase enzyme and said second DNA recombinase enzyme carrying said at least one mutation in their catalytic site when co-expressed and forming a complex show the catalytic activity of a DNA recombinase. The invention further relates to nucleic acid molecules encoding said genetically engineered DNA recombinases and complexes, as well as to pharmaceutical compositions comprising the same. The invention further provides the use of said genetically engineered DNA recombinases, complexes, nucleic acid molecules and pharmaceutical compositions in medicine and specifically for treating genetic disorders such as hemophilia A.