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PLoS One. 2014 Dec 1;9(12):e113729. doi: 10.1371/journal.pone.0113729. eCollection 2014.

Selection of intracellular single-domain antibodies targeting the HIV-1 Vpr protein by cytoplasmic yeast two-hybrid system.

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Inserm U1068, CRCM, Marseille, France; Institut Paoli-Calmettes, Marseille, France; Aix-Marseille Université UM105, Marseille, France; CNRS, UMR7258, CRCM, Marseille, France.
Institut Cochin, CNRS UMR8104, Paris, France; Université Paris Descartes, Paris, France; Inserm U1016, Paris, France.


The targeting of HIV-1 using antibodies is of high interest as molecular tools to better understand the biology of the virus or as a first step toward the design of new inhibitors targeting critical viral intracellular proteins. Small and highly stable llama-derived single-domain antibodies can often be functionally expressed as intracellular antibodies in the cytoplasm of eukaryotic cells. Using a selection method based on the Sos Recruitment System, a cytoplasmic yeast two-hybrid approach, we have isolated single-domain antibodies able to bind HIV-1 Vpr and Capside proteins in the yeast cytoplasm. One anti-Vpr single domain antibody was able to bind the HIV-1 regulatory Vpr protein in the cytoplasm of eukaryotic cells, leading to its delocalization from the nucleus to the cytoplasm. To our knowledge, this is the first description of a functional single-domain intrabody targeting HIV-1 Vpr, isolated using an in vivo cytoplasmic selection method that alleviates some limitations of the conventional yeast two-hybrid system.

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