Technische Universitát Dresden

Max-Planck-Gesellschaft Zur F&#xf6;rderung Der Wissenschaften E. V.

Heinrich-Pette-Institut Leibniz-Institut Für Experimentelle Virologie-Stiftung Bürgerlichen Rechts

Heinrich-Pette-Institut Für Experimentelle Virologie Und Immunologie an Der Universität Hamburg

Heinrich-Pette-Institut, Leibniz-Institut Fur Experimentelle Virologie

Frank Buchholz

Madina Karimova

Joachim Hauber

A Francis Stewart

Jan Chemnitz

Indrani Sarkar

Janet Karpinski

Martin Schneider

Felix Lansing

Ilona Hauber

Janet Chusainow

Francis A Stewart

The invention provides methods 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).

Methods and means for genetic alteration of genomes utilizing designer DNA recombining enzymes

The present invention relates to a method for preparing an expression vector encoding a tailored recombinase, which tailored recombinase is capable of recombining asymmetric target sequences within the long terminal repeat (LTR) of proviral DNA of a plurality of retrovirus strains inserted into the genome of a host cell, as well as to the obtained expression vector, cells transfected with this, expressed recombinase and pharmaceutical compositions comprising the expression vector, cells and/or recombinase. Pharmaceutical compositions are useful, e.g., in treatment and/or prevention of retrovirus infection. In particular, asymmetric target sequences present in a plurality of HIV strains are disclosed, as well as tailored recombinases capable of combining these sequences (Tre 3.0 and 4.0) and expression vectors encoding them.

Tailored recombinase for recombining asymmetric target sites in a plurality of retrovirus strains

The invention relates to the use of a protein with recombinase activity to catalyze a site-specific DNA recombination and a method for producing a site-specific DNA recombination. The invention is applicable alone or in combination with other recombinase systems for genetic manipulation, for example in medical research. The objective of the invention is solved by the use of a protein with recombinase activity to catalyze a site-specific DNA recombination at, preferably at two, recognition sites that are identical or reverse complementary to each other. The invention also includes a method for producing a site-specific DNA recombination comprising the steps of a) providing a cell comprising at least two recognition sites that are identical or reverse complementary to each other; and b) contacting a protein with recombinase activity with the recognition sites, thereby producing the site-specific DNA-recombination.

Protein with recombinase activity for site-specific DNA-recombination

The present invention relates to a method for preparing an expression vector encoding a well-tolerated and highly specific tailored recombinase, which tailored recombinase is capable of recombining asymmetric target sequences within the long terminal repeat (LTR) of proviral DNA of a plurality of retrovirus strains which may be inserted into the genome of a host cell, as well as to the obtained expression vector, cells transfected with these, expressed recombinase and pharmaceutical compositions comprising the expression vector, cells and/or recombinase. Pharmaceutical compositions are useful, e.g., in treatment and/or prevention of retrovirus infection, in particular, HIV infection. In particular, the invention relates to well-tolerated and highly specific tailored recombinases capable of combining asymmetric target sequences in a more than 90% of HIV-strains, thereby excising the HIV-1 sequences, and expression vectors encoding them.

Well-tolerated and highly specific tailored recombinase for recombining asymmetric target sites in a plurality of retrovirus strains

The present invention is directed to a method for preparing an expression vector encoding a tailored recombinase, wherein said tailored recombinase recombines asymmetric target sites within the LTR of proviral DNA of a retrovirus inserted into the genome of a host cell and is useful as means for excising the provirus from the genome of the host cell. The present invention further relates to an in vitro-method of optimising the treatment of a retroviral infection of a subject and to the use of tailored recombinases for the preparation of pharmaceutical compositions for reducing the viral load in a subjected infected by a retrovirus.

Use of tailored recombinases for the treatment of retroviral infections

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Dresden, Germany

Frank Buchholz