Radosław Kamil Ejsmont

Laboratory of Neurogenetics, VIB / K.U.Leuven

Campus Gasthuisberg - O&N4 07.05
Herestraat 49 - bus 602
B-3000 Leuven, Belgium

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Research focus

Systematic generation of live gene expression reporters
(FlyFos project)

Radosław Kamil Ejsmont, Maria Bogdanzaliewa, Pavel Tomančák
Tomančák Lab, MPI-CBG Dresden

In order to visualize gene expression patterns in live embryos, we constructed two complementary genomic fosmid libraries for Drosophila melanogaster and Drosophila pseudoobscura that permit seamless modification of large genomic clones by high-throughput recombineering and direct transgenesis. The fosmid transgenes rescue mutant phenotypes, recapitulate endogenous gene expression patterns and in some cases allow imaging of gene products in living animals. The D.pseudoobscura transgenes rescue RNAi phenotypes when introduced into the D.melanogaster genome, providing a convenient control for the specificity of the knockdown. These libraries will, in combination with recombineering technology, enable systematic analysis and manipulation of gene activity across species.

We are currently testing various tags suitable for live imaging. We are developing a universal tagging system that enables exchange of tags in vivo without the time-consuming transgenesis. Finally, we are optimizing a HT liquid-culture recombineering pipeline to introduce deletions into the already tagged fosmid constructs to dissect cis-regulatory sequences.

We will leverage the astonishing efficiency of our toolkit to establish a genome-wide resource of tagged fosmid transgenes and organize a distributed, community-driven transgenesis effort. We will use the toolkit to test specific hypothesis of sequence determinants of gene expression pattern divergence.