RT Journal Article SR Electronic T1 Protein manipulation using single copies of short peptide tags in cultured cells and in Drosophila melanogaster JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.04.06.027599 DO 10.1101/2020.04.06.027599 A1 M. Alessandra Vigano A1 Clara-Maria Ell A1 Manuela MM Kustermann A1 Gustavo Aguilar A1 Shinya Matsuda A1 Ning Zhao A1 Timothy J Stasevich A1 George Pyrowolakis A1 Markus Affolter YR 2020 UL http://biorxiv.org/content/early/2020/05/05/2020.04.06.027599.abstract AB Cellular development and specialized cellular functions are regulated processes which rely on highly dynamic molecular interactions among proteins, distributed in all cell compartments. Analysis of these interactions and their mechanisms of action has been one of the main topics in cellular and developmental research over the last fifty years. Studying and understanding the functions of proteins of interest (POIs) has been mostly achieved by their alteration at the genetic level and the analysis of the phenotypic changes generated by these alterations. Although genetic and reverse genetic technologies contributed to the vast majority of information and knowledge we have gathered so far, targeting specific interactions of POIs in a time- and space-controlled manner or analyzing the role of POIs in dynamic cellular processes such as cell migration or cell division would require more direct approaches. The recent development of specific protein binders, which can be expressed and function intracellularly, together with several improvements in synthetic biology techniques, have contributed to the creation of a new toolbox for direct protein manipulations. We selected a number of short tag epitopes for which protein binders from different scaffolds have been developed and tested whether these tags can be bound by the corresponding protein binders in living cells when they are inserted in a single copy in a POI. We indeed find that in all cases, a single copy of a short tag allows protein binding and manipulation. Using Drosophila, we also find that single short tags can be recognized and allow degradation and relocalization of POIs in vivo.Competing Interest StatementThe authors have declared no competing interest.