Journal Article

Generation of a versatile BiFC ORFeome library for analyzing protein-protein interactions in live Drosophila

Giraud, Guillaume, Institut de Génomique Fonctionnelle de Lyon (IGFL) ; École normale supérieure de Lyon (ENS de Lyon) ; Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL) ; Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), Spinelli, Lionel, Universität Zürich [Zürich] = University of Zurich (UZH), Brun, Christine, Ahmed, Alexis, Ahr, Damien, Furger, Edy, Basler, Konrad, Zhejiang University [Hangzhou, China], Vanderperre, Solene, Merabet, Samir, Ajuria, Leiore, Bischof, Johannes, Theories and Approaches of Genomic Complexity (TAGC) ; Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Duffraisse, Marilyne, Paul, Rachel, Björklund, Mikael

2018

Overview

Transcription factors achieve specificity by establishing intricate interaction networks that will change depending on the cell context. Capturing these interactions in live condition is however a challenging issue that requires sensitive and non-invasive methods. We present a set of fly lines, called 'multicolor BiFC library', which covers most of the Drosophila transcription factors for performing Bimolecular Fluorescence Complementation (BiFC). The multicolor BiFC library can be used to probe two different binary interactions simultaneously and is compatible for large-scale interaction screens. The library can also be coupled with established Drosophila genetic resources to analyze interactions in the developmentally relevant expression domain of each protein partner. We provide proof of principle experiments of these various applications, using Hox proteins in the live Drosophila embryo as a case study. Overall this novel collection of ready-to-use fly lines constitutes an unprecedented genetic toolbox for the identification and analysis of protein-protein interactions in vivo.

• Publisher: CCSD,eLife Sciences Publication,eLife Science Publications, Ltd,eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
• Subject: Animals / Animals, Genetically Modified / Biochemistry, Molecular Biology / Color / Complementation / Development Biology / DNA binding proteins / Drosophila / Drosophila - embryology / Drosophila - genetics / Drosophila - metabolism / Drosophila Proteins - genetics / Drosophila Proteins - metabolism / Embryo / Embryo, Nonmammalian - embryology / Embryo, Nonmammalian - metabolism / Embryos / Experiments / Fluorescence / Gene Expression Regulation, Developmental / Gene Library / Genetic analysis / Genetic aspects / Genetic research / Genetic resources / Genetics and Genomics / Hox / Insects / Library collections / Life Sciences / Luminescent Proteins - chemistry / Luminescent Proteins - genetics / Luminescent Proteins - metabolism / Microscopy, Fluorescence - methods / Molecular biology / Molecular Networks / Novels / Protein Binding / Protein expression / Protein interaction / Protein Interaction Mapping - methods / Protein-protein interactions / Proteins / Tools and Resources / Transcription factors / Transcription Factors - genetics / Transcription Factors - metabolism