SITE-SPECIFIC RECOMBINASES FOR EFFICIENT AND SPECIFIC GENOME EDITING

The invention relates generally to the field of genome editing and provides a method of generating DNA recombinases and provides DNA recombinases, which efficiently and specifically recombine genomic target sequences as obligate DNA recombinase enzymes. More specifically, the invention provides genetically engineered DNA recombinases for efficient and specific genome editing, comprising an obligate complex of recombinases, wherein said complex comprises at least a first recombinase enzyme and at least 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 a catalytic site, wherein said first recombinase enzyme and said second recombinase enzyme carrying said at last one mutation in a catalytic site, when expressed in isolation, do not show the catalytic activity of a DNA recombinase; and wherein the catalytic activity as a DNA recombinase in said obligate complex of recombinases is complemented. The invention further relates to nucleic acid molecules encoding said genetically engineered DNA recombinases and obligate complexes, as well as to the use of said genetically engineered DNA recombinases and obligate complexes and nucleic acid molecules in a pharmaceutical composition.