title: A versatile in vivo system for directed dissection of gene expression patterns.
abstract: Tissue-specific gene expression using the upstream activating sequence (UAS)–GAL4 binary system has facilitated genetic dissection of many biological processes in Drosophila melanogaster. Refining GAL4 expression patterns or independently manipulating multiple cell populations using additional binary systems are common experimental goals. To simplify these processes, we developed a convertible genetic platform, the integrase swappable in vivo targeting element (InSITE) system. This approach allows GAL4 to be replaced with any other sequence, placing different genetic effectors under the control of the same regulatory elements. Using InSITE, GAL4 can be replaced with LexA or QF, allowing an expression pattern to be repurposed. GAL4 can also be replaced with GAL80 or split-GAL4 hemi-drivers, allowing intersectional approaches to refine expression patterns. The exchanges occur through efficient in vivo manipulations, making it possible to generate many swaps in parallel. This system is modular, allowing future genetic tools to be easily incorporated into the existing framework.
authors.affiliation: Department of Neurobiology, Stanford University, Stanford, California, USA.
authors.forename: Daryl M
authors.initials: DM
authors.lastname: Gohl
authors.forename: Marion A
authors.initials: MA
authors.lastname: Silies
authors.forename: Xiaojing J
authors.initials: XJ
authors.lastname: Gao
authors.forename: Sheetal
authors.initials: S
authors.lastname: Bhalerao
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Luongo
authors.forename: Chun-Chieh
authors.initials: CC
authors.lastname: Lin
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Potter
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Clandinin
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D012097:Repressor Proteins / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D000099160:LexA Repressor Protein
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases; D014157:Transcription Factors; D000099160:LexA Repressor Protein; C052130:GAL4 protein, S cerevisiae; C055002:GAL80 protein, S cerevisiae