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Targeted In Vivo Inhibition of Specific Protein–Protein Interactions Using Recombinant Antibodies

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ZÁBRADY Matej HRDINOVÁ Vendula MÜLLER Bruno CONRAD Udo HEJÁTKO Jan JANDA Lubomír

Rok publikování 2014
Druh Článek v odborném periodiku
Časopis / Zdroj PLOS ONE : Public Library of Science
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
www http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0109875
Doi http://dx.doi.org/10.1371/journal.pone.0109875
Obor Genetika a molekulární biologie
Klíčová slova recombinant antibodies;immunomodulation;inhibition;protein-protein interactions;cytokinin signaling pathway
Popis With the growing availability of genomic sequence information, there is an increasing need for gene function analysis. Antibody-mediated “silencing” represents an intriguing alternative for the precise inhibition of a particular function of biomolecules. Here, we describe a method for selecting recombinant antibodies with a specific purpose in mind, which is to inhibit intrinsic protein–protein interactions in the cytosol of plant cells. Experimental procedures were designed for conveniently evaluating desired properties of recombinant antibodies in consecutive steps. Our selection method was successfully used to develop a recombinant antibody inhibiting the interaction of ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER PROTEIN 3 with such of its upstream interaction partners as the receiver domain of CYTOKININ INDEPENDENT HISTIDINE KINASE 1. The specific down-regulation of the cytokinin signaling pathway in vivo demonstrates the validity of our approach. This selection method can serve as a prototype for developing unique recombinant antibodies able to interfere with virtually any biomolecule in the living cell.
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