Refinement of tools for targeted gene expression in Drosophila.
A wide variety of biological experiments rely on the ability to express an exogenous gene in a transgenic animal at a defined level and in a spatially and temporally controlled pattern. We describe major improvements of the methods available for achieving this objective in Drosophila melanogaster. We have systematically varied core promoters, UTRs, operator sequences, and transcriptional activating domains used to direct gene expression with the GAL4, LexA, and Split GAL4 transcription factors and the GAL80 transcriptional repressor. The use of site-specific integration allowed us to make quantitative comparisons between different constructs inserted at the same genomic location. We also characterized a set of PhiC31 integration sites for their ability to support transgene expression of both drivers and responders in the nervous system. The increased strength and reliability of these optimized reagents overcome many of the previous limitations of these methods and will facilitate genetic manipulations of greater complexity and sophistication.
Pfeiffer
Barret D
BD
Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. pfeifferb@janelia.hhmi.org
Ngo
Teri-T B
TT
Hibbard
Karen L
KL
Murphy
Christine
C
Jenett
Arnim
A
Truman
James W
JW
Rubin
Gerald M
GM
D000818:Animals; D030801:Animals, Genetically Modified; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005818:Genetic Engineering; D005819:Genetic Markers; D005821:Genetic Techniques; D005822:Genetic Vectors; D012045:Regulatory Sequences, Nucleic Acid; D020133:Reverse Transcriptase Polymerase Chain Reaction; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D019076:Transgenes
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005819:Genetic Markers; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors