1030C Poster - 16. Techniques and technology
Saturday April 09, 1:30 PM - 3:30 PM

A toolkit to generate interconvertible overexpression Drosophila transgenes


Authors:
Luis Alberto Baena Lopez 1; Franz Wendler 1; Sangbin Park 2; Claire Hill 1; Alessia Galasso 1; Kathleen Chang 2; Iman Awan 1; Yulia Sudarikova 1; Mar Bustamante-Sequeiros 1; Sichen Liu 1; Ethan Seung 1; Gabrielle Aisabonoko 1; Seung Kim 2

Affiliations:
1) University of Oxford; 2) Department of Developmental Biology, Stanford University School of Medicine

Keywords:
q. other (Ectopic Gene activation); f. classroom undergraduate research experience (CURE)

Three independent binary systems for conditional gene expression (Gal4/UAS; LexA/LexAop; QF/QUAST) have greatly aided the genetic investigations using Drosophila as a model organism. However, each of these inducible gene expression systems requires the independent generation of non-interchangeable fly repositories. To alleviate this important logistic disadvantage and the need of triplicating transgene repositories for combinatorial experiments, we developed a modular vector that contains the regulatory elements for all three binary systems. This enables the separate or simultaneous overexpression of new transgenes through any of the available transcriptional factors (Gal4-, LexA- or QF), thus offering new experimental possibilities for in vivo cDNA overexpression (robust RNAi downregulation needs to be individually tested). Additionally, each transcriptional regulatory element (UAS, LexAop, QUAS) was flanked by two different but compatible recombination sites that facilitate their elimination from the genome with spatial and temporal control upon exposure to adequate recombinases. This allows the efficient generation of genetic mosaics within somatic cells as well as the elimination in the germline of specific gene-activating sequences. These unique features of our genetic toolkit provide unprecedented experimental possibilities and logistical advantages such as the generation of new fly lines with a subset of gene regulatory elements of interest upon excising the unwanted in a common precursor strain containing all of them.