公开(公告)号：US20210147878A1

公开(公告)日：2021-05-20

申请号：US16622937

申请日：2018-06-14

Applicant: TECHNISCHE UNIVERSITÄT DRESDEN

Inventor： Frank BUCHHOLZ ,  Martin SCHNEIDER ,  Felix LANSING

IPC: C12N15/90 ,  C12Q1/6869 ,  C12N15/10 ,  C12N9/22

Abstract: The invention provides methods and means for specifically altering the DNA sequence in a genome, in particular for genome editing by deleting or replacing a sequence of interest. Advantageously, the invention uses two non-identical sequences naturally occurring in a genome as target sites two which DNA-recombining enzymes are generated. The invention is in particular useful for medicine, in particular to repair a mutation in a genome or to delete predefined genetic material from cells or tissue and to cure diseases. An advantage of the invention is that it allows precise site directed altering of DNA without engaging host DNA repair pathways and thereby works without inducing random insertions and deletions (indels).

公开(公告)号：US20230235364A1

公开(公告)日：2023-07-27

申请号：US18046066

申请日：2022-10-12

Applicant: TECHNISCHE UNIVERSITÄT DRESDEN

Inventor： Frank BUCHHOLZ ,  Martin SCHNEIDER ,  Felix LANSING

IPC: C12N15/90 ,  C12N9/22 ,  C12N15/10 ,  C12Q1/6869

CPC classification number: C12N15/907 ,  C12N9/22 ,  C12N15/1058 ,  C12Q1/6869

Abstract: The invention provides methods and means for specifically altering the DNA sequence in a genome, in particular for genome editing by deleting or replacing a sequence of interest. Advantageously, the invention uses two non-identical sequences naturally occurring in a genome as target sites two which DNA-recombining enzymes are generated. The invention is in particular useful for medicine, in particular to repair a mutation in a genome or to delete predefined genetic material from cells or tissue and to cure diseases. An advantage of the invention is that it allows precise site directed altering of DNA without engaging host DNA repair pathways and thereby works without inducing random insertions and deletions (in-dels).

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.