A toolkit to wire synthetic transcriptional circuits in Drosophila melanogaster
Authors: Aya Gomaa 1,2,3; Ariane Ramaekers 3; Radoslaw Ejsmont 2
Affiliations: 1) University of Paris, Paris.; 2) Le Centre de recherches interdisciplinaires, Paris.; 3) Curie Institute, Paris.
Keywords: a. core promoters and general transcription factors; q. other (Synthetic biology)
Development is an extraordinarily complex and tightly regulated process. A significant part of developmental regulation depends on gene regulatory networks (GRNs), consisting of interacting transcription factors governing spatial and temporal gene expression. Synthetic biology opens a new way to study GRNs: on top of deciphering natural GRNs, scientists reconstruct synthetic ones using engineered transcription factors and target genes. Existing synthetic GRNs have primarily been established in prokaryotes that benefit from toolkits consisting of ‘easy to assemble’ well-studied genetic elements, such as promoters and transcription factors. In our project, we are developing a toolkit to reconstruct synthetic GRNs in vivo, in Drosophila melanogaster (D.mel). Our toolkit enables massive combinatorial assembly of synthetic transcription factors (sTFs) using a library of DNA binding domains, activating or repressing domains and a collection of engineered cis-regulatory binding sites that are absent from the (D.mel) genome. We are establishing the gene expression profiles derived from these sTFs individually or in a variety of combinations. Ultimately, sTFs will be used to recreate motifs that occur naturally in GRNs and logic gates. By providing this sTFs library we hope to facilitate the implementation of synthetic GRNs in D.mel and other model organisms, thus enabling in vivo modeling of the native developmental networks.