title: Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity.
abstract: Current combination antiretroviral therapies (cART) efficiently suppress HIV-1 reproduction in humans, but the virus persists as integrated proviral reservoirs in small numbers of cells. To generate an antiviral agent capable of eradicating the provirus from infected cells, we employed 145 cycles of substrate-linked directed evolution to evolve a recombinase (Brec1) that site-specifically recognizes a 34-bp sequence present in the long terminal repeats (LTRs) of the majority of the clinically relevant HIV-1 strains and subtypes. Brec1 efficiently, precisely and safely removes the integrated provirus from infected cells and is efficacious on clinical HIV-1 isolates in vitro and in vivo, including in mice humanized with patient-derived cells. Our data suggest that Brec1 has potential for clinical application as a curative HIV-1 therapy.
authors.affiliation: Medical Systems Biology, UCC, Medical Faculty Carl Gustav Carus, TU Dresden, Germany.|Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Janet
authors.initials: J
authors.lastname: Karpinski
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Ilona
authors.initials: I
authors.lastname: Hauber
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Chemnitz
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site Hamburg, Germany.
authors.forename: Carola
authors.initials: C
authors.lastname: Schäfer
authors.affiliation: Medical Systems Biology, UCC, Medical Faculty Carl Gustav Carus, TU Dresden, Germany.
authors.forename: Maciej
authors.initials: M
authors.lastname: Paszkowski-Rogacz
authors.affiliation: Medical Systems Biology, UCC, Medical Faculty Carl Gustav Carus, TU Dresden, Germany.
authors.forename: Deboyoti
authors.identifier: http://orcid.org/0000-0002-2300-4333
authors.initials: D
authors.lastname: Chakraborty
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Niklas
authors.initials: N
authors.lastname: Beschorner
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site Hamburg, Germany.
authors.forename: Helga
authors.initials: H
authors.lastname: Hofmann-Sieber
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site Hamburg, Germany.|Department of Anesthesiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Ulrike C
authors.initials: UC
authors.lastname: Lange
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site Hamburg, Germany.
authors.forename: Adam
authors.initials: A
authors.lastname: Grundhoff
authors.affiliation: Institute for Clinical Genetics, University Hospital and Medical Faculty Carl Gustav Carus, TU Dresden, Germany.
authors.forename: Karl
authors.initials: K
authors.lastname: Hackmann
authors.affiliation: Institute for Clinical Genetics, University Hospital and Medical Faculty Carl Gustav Carus, TU Dresden, Germany.
authors.forename: Evelin
authors.initials: E
authors.lastname: Schrock
authors.affiliation: Structural Bioinformatics, BIOTEC TU Dresden, Germany.
authors.forename: Josephine
authors.initials: J
authors.lastname: Abi-Ghanem
authors.affiliation: Structural Bioinformatics, BIOTEC TU Dresden, Germany.
authors.forename: M Teresa
authors.initials: MT
authors.lastname: Pisabarro
authors.affiliation: Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany.
authors.forename: Vineeth
authors.initials: V
authors.lastname: Surendranath
authors.affiliation: Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Schambach
authors.affiliation: Agaplesion Diakonieklinikum, Gynecology, Hamburg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Lindner
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site Hamburg, Germany.|Infectious Diseases Unit, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: van Lunzen
authors.affiliation: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site Hamburg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Hauber
authors.affiliation: Medical Systems Biology, UCC, Medical Faculty Carl Gustav Carus, TU Dresden, Germany.|Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Buchholz
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D002478:Cells, Cultured; D019020:Directed Molecular Evolution / Q000379:methods*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011533:Proviruses / Q000187:drug effects* / Q000235:genetics; D045522:Recombinases / Q000378:metabolism / Q000494:pharmacology*; D016662:Virus Integration / Q000187:drug effects* / Q000235:genetics
chemical_list: D000998:Antiviral Agents; D045522:Recombinases