A Flippase-Mediated GAL80/GAL4 Intersectional Resource for Dissecting Appendage Development in Drosophila.
Drosophila imaginal discs provide an ideal model to study processes important for cell signaling and cell specification, tissue differentiation, and cell competition during development. One challenge to understanding genetic control of cellular processes and cell interactions is the difficulty in effectively targeting a defined subset of cells in developing tissues in gene manipulation experiments. A recently developed Flippase-induced intersectional GAL80/GAL4 repression method incorporates several gene manipulation technologies in Drosophila to enable such fine-scale dissection in neural tissues. In particular, this approach brings together existing GAL4 transgenes, newly developed enhancer-trap flippase transgenes, and GAL80 transgenes flanked by Flippase recognition target sites. The combination of these tools enables gene activation/repression in particular subsets of cells within a GAL4 expression pattern. Here, we expand the utility of a large collection of these enhancer-trap flippase transgenic insertion lines by characterizing their expression patterns in third larval instar imaginal discs. We screened 521 different enhancer-trap flippase lines and identified 28 that are expressed in imaginal tissues, including two transgenes that show sex-specific expression patterns. Using a line that expresses Flippase in the wing imaginal disc, we demonstrate the utility of this intersectional approach for studying development by knocking down gene expression of a key member of the planar cell polarity pathway. The results of our experiments show that these enhancer-trap flippase lines enable fine-scale manipulation in imaginal discs.
Department of Biology, University of Oklahoma, Norman, Oklahoma 73019.
Brittany N
BN
Smith
Department of Biology, University of Oklahoma, Norman, Oklahoma 73019.
Arash M
AM
Ghazanfari
Department of Biology, University of Oklahoma, Norman, Oklahoma 73019 Department of Biological and Health Sciences, Texas A&M University, Kingsville, Texas 78363.
Rudolf A
RA
Bohm
Department of Biology, University of Oklahoma, Norman, Oklahoma 73019.
William P
WP
Welch
Department of Biology, University of Oklahoma, Norman, Oklahoma 73019 Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211.
Bing
B
Zhang
Department of Biology, University of Oklahoma, Norman, Oklahoma 73019 masly@ou.edu.
John P
http://orcid.org/0000-0003-3448-0914
JP
Masly
D000818:Animals; D004330:Drosophila / Q000196:embryology / Q000235:genetics* / Q000378:metabolism*; D029721:Drosophila Proteins / Q000378:metabolism*; D015870:Gene Expression; D005786:Gene Expression Regulation; D060227:Imaginal Discs / Q000196:embryology / Q000378:metabolism; D007814:Larva; D011995:Recombination, Genetic; D012097:Repressor Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; D012097:Repressor Proteins; D014157:Transcription Factors
Drosophila; enhancer-trap Flippase; genetic manipulation; imaginal discs