Tailored recombinase for recombining asymmetric target sites in a plurality of retrovirus strains
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.
1. An evolution vector comprising a nucleic acid selected from the group consisting of SEQ ID NOS: 1, 2, and 8-15, wherein the evolution vector further encodes a recombinase, and wherein expression of the recombinase is controlled by an inducible promoter.
2. The evolution vector of claim 1 , comprising the nucleic acid of SEQ ID NO: 1.
3. The evolution vector of claim 1 , comprising the nucleic acid of SEQ ID NO: 2.
4. The evolution vector of claim 1 , wherein the evolution vector is a plasmid.
5. The evolution vector of claim 1 , wherein the vector is capable of being recombined by a tailored recombinase comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 40-64.
6. The evolution vector of claim 1 , wherein the recombinase is selected from the group comprising Cre, Tre, Dre, Zre and Shew.
7. The evolution vector of claim 1 , wherein the recombinase is a randomly mutated recombinase.