Reverse genetics in <i>drosophila</i>: From sequence to phenotype using <i>UAS</i>‐RNAi transgenic flies

Enerly, Espen; Larsson, Jonas; Lambertsson, Andrew

doi:10.1002/gene.10111

Cited by 12 publications

(9 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…DNA from coding sequence of the tenectin gene was amplified by PCR and cloned into the pUAST vector as an inverted repeat as described (Enerly et al, 2002; Kennerdell and Carthew, 2000). Amplification used sense and antisense primers containing Eco RI and Xba I sites and Blg II and Xho I sites and were 5’-CCGGAATTCTGTTGAAATCGACACGAAGC-3’, 5’-GAAGATCTTACCTCAGGCTCCTCATGCT-3’, 5’-GCTCTAGATGTTGAAATCGACACGAAG-3’ and 5’-CCGCTCGAGTACCTCAGGCTCCTCATGCT-3’ respectively.…”

Section: Methodsmentioning

confidence: 99%

Tenectin is a novel αPS2βPS integrin ligand required for wing morphogenesis and male genital looping in Drosophila

Fraichard

Bougé

Kendall

et al. 2010

Developmental Biology

View full text Add to dashboard Cite

Morphogenesis of the adult structures of holometabolous insects is regulated by ecdysteroids and juvenile hormones and involves cell-cell interactions mediated in part by the cell surface integrin receptors and their extracellular matrix (ECM) ligands. These adhesion molecules and their regulation by hormones are not well characterized. We describe the gene structure of a newly described ECM molecule, tenectin, and demonstrate that it is a hormonally regulated ECM protein required for proper morphogenesis of the adult wing and male genitalia. Tenectin’s function as a new ligand of the PS2 integrins is demonstrated by both genetic interactions in the fly and by cell spreading and cell adhesion assays in cultured cells. Its interaction with the PS2 integrins is dependent on RGD and RGD-like motifs. Tenectin’s function in looping morphogenesis in the development of the male genitalia led to experiments that demonstrate a role for PS integrins in the execution of left-right asymmetry.

show abstract

Section: Methodsmentioning

confidence: 99%

Tenectin is a novel αPS2βPS integrin ligand required for wing morphogenesis and male genital looping in Drosophila

Fraichard

Bougé

Kendall

et al. 2010

Developmental Biology

View full text Add to dashboard Cite

show abstract

“…The evidence supporting cell autonomy for PTGS in Drosophila came initially from a failure to see spreading of the phenotypic effects outside of regions defined by Gal4 expression (e.g. (56,(61)(62)(63) Van Roessel et al (64) ) found, by examining the PTGS of GFP variants using laser scanning confocal microscopy, that the effect could be strictly limited to engrailed defined embryonic segments. An important point from this study was that GAP junctions present between epidermal and ectodermal cells would not support PTGS spreading.…”

Section: Reduction-of-function Transgenicsmentioning

confidence: 98%

The genetics of Drosophila transgenics

Roman¹

2004

BioEssays

View full text Add to dashboard Cite

In Drosophila, the genetic approach is still the method of choice for answering fundamental questions on cell biology, signal transduction, development, physiology and behavior. In this approach, a gene's function is ascertained by altering either the amount or quality of the gene product, and then observing the consequences. The genetic approach is itself polymorphous, encompassing new and more complex techniques that typically employ the growing collections of transgenes. The keystone of these modern Drosophila transgenic techniques has been the Gal4 binary system. Recently, several new techniques have modified this binary system to offer greater control over the timing, tissue specificity and magnitude of gene expression. Additionally, the advances in post-transcriptional gene silencing, or RNAi, have greatly expanded the ability to knockdown almost any gene's function. Regardless of the growing experimental intricacy, the application of these advances to modify gene activity still obeys the fundamental principles of genetic analysis. Several of these transgenic techniques, which offer more precise control over a gene's activity, will be reviewed here with a discussion on how they may be used for determining a gene's function.

show abstract

“…Interference RNA against NELF-E was shown recently to cause developmental defects or lethality depending on where the interference RNA was expressed (Enerly et al 2002). For our experiment, one transgenic fly line, 17A10, contained a transgene encoding an interference RNA against NELF-E that was controlled by Gal4p.…”

Section: Depletion Of Nelf-e In Salivary Glands Reduces the Level Of mentioning

confidence: 99%

“…The fly line 17A10 containing a NELF-E RNAi construct controlled by the Gal4 UAS has been described previously (Enerly et al 2002). To direct expression of RNAi in salivary glands, the NELF-E RNAi flies were mated to the Bloomington stock #1824 [genotype: y1 w*; P(w +mW.hs = GawB)AB1] that expresses Gal4p in salivary glands.…”

Section: Permanganate Genomic Footprintingmentioning

confidence: 99%

NELF and DSIF cause promoter proximal pausing on thehsp70promoter inDrosophila

et al. 2003

Self Cite

View full text Add to dashboard Cite

NELF and DSIF collaborate to inhibit elongation by RNA polymerase IIa in extracts from human cells. A multifaceted approach was taken to investigate the potential role of these factors in promoter proximal pausing on the hsp70 gene in Drosophila. Immunodepletion of DSIF from a Drosophila nuclear extract reduced the level of polymerase that paused in the promoter proximal region of hsp70. Depletion of one NELF subunit in salivary glands using RNA interference also reduced the level of paused polymerase. In vivo protein-DNA cross-linking showed that NELF and DSIF associate with the promoter region before heat shock. Immunofluorescence analysis of polytene chromosomes corroborated the cross-linking result and showed that NELF, DSIF, and RNA polymerase IIa colocalize at the hsp70 genes, small heat shock genes, and many other chromosomal locations. Finally, following heat shock induction, DSIF and polymerase but not NELF were strongly recruited to chromosomal puffs harboring the hsp70 genes. We propose that NELF and DSIF cause polymerase to pause in the promoter proximal region of hsp70. The transcriptional activator, HSF, might cause NELF to dissociate from the elongation complex. DSIF continues to associate with the elongation complex and could serve a positive role in elongation.

show abstract

Reverse genetics in drosophila: From sequence to phenotype using UAS‐RNAi transgenic flies

Cited by 12 publications

References 18 publications

Tenectin is a novel αPS2βPS integrin ligand required for wing morphogenesis and male genital looping in Drosophila

Tenectin is a novel αPS2βPS integrin ligand required for wing morphogenesis and male genital looping in Drosophila

The genetics of Drosophila transgenics

NELF and DSIF cause promoter proximal pausing on thehsp70promoter inDrosophila

Contact Info

Product

Resources

About