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  • DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as an Alk target in the Drosophila visceral mesoderm.

DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as an Alk target in the Drosophila visceral mesoderm.

Development (Cambridge, England) (2021-12-15)
Patricia Mendoza-Garcia, Swaraj Basu, Sanjay Kumar Sukumar, Badrul Arefin, Georg Wolfstetter, Vimala Anthonydhason, Linnea Molander, Ezgi Uçkun, Henrik Lindehell, Cristina Lebrero-Fernandez, Jan Larsson, Erik Larsson, Mats Bemark, Ruth H Palmer
ABSTRACT

Development of the Drosophila visceral muscle depends on Anaplastic Lymphoma Kinase (Alk) receptor tyrosine kinase (RTK) signaling, which specifies founder cells (FCs) in the circular visceral mesoderm (VM). Although Alk activation by its ligand Jelly Belly (Jeb) is well characterized, few target molecules have been identified. Here, we used targeted DamID (TaDa) to identify Alk targets in embryos overexpressing Jeb versus embryos with abrogated Alk activity, revealing differentially expressed genes, including the Snail/Scratch family transcription factor Kahuli (Kah). We confirmed Kah mRNA and protein expression in the VM, and identified midgut constriction defects in Kah mutants similar to those of pointed (pnt). ChIP and RNA-Seq data analysis defined a Kah target-binding site similar to that of Snail, and identified a set of common target genes putatively regulated by Kah and Pnt during midgut constriction. Taken together, we report a rich dataset of Alk-responsive loci in the embryonic VM and functionally characterize the role of Kah in the regulation of embryonic midgut morphogenesis.

MATERIALS
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Product Description

Roche
SP6/T7 Transcription Kit, sufficient for 2 x 20 assays (stadard transcription), kit of 1 (12 components), suitable for DNA sequencing, suitable for hybridization