Techniques Platform.
Perrimon group/ RNAi centers
Cell based RNAi screening. Increase in imaging screens (from plate-reader screens).
1-3 million images per std experiment, taken an automatically analyzed.
2-10Tb per screen, currently 35Tb of data. Trying to move to in vivo whole embryos(?)
Neuron cell biology
clone many genes in parallel with BAC recombineering with flanking FRT sites.
Generate 30 modified BACs in parallel using FLP
about half rabs are neuron specific. Approach with FRT can be used for targeted gene knockout after characterization.
Virtual Fly Brain
hard to reconstruct maps from literature.
Different naming conventions in literature – different terms for the same thing ame terms for slightly different things.
Solution: online interactive database: virtual fly brain.
Standard ontology. Open source software and display. 130,000 phenotypes, 30,000 patterns. Integrated to flybase.
Proteome scale protein interaction map. DPiM
(Spyros Artavanis-Tsakonas, Harvard, consortium pipeline) S2-cells to mass-spec. Data on flybase (straight). Scores also prey-prey interactions.
‘we have done expensive bioninformatincs analysis on the network’.
High quality protein map. Looks pretty convincing, sparser than Y2H, complete with few novel elements, find functional, transient complex-complex interactions.
Enhancer-trap Flippase Gal4/Gal80 ‘FINGR method’
most gal4 lines are leaky. 1 method, split gal4 binding and activating domain to get more faithful expression. Not compatable with previous gal4s. Gal80 less available.
Express flipase in a tissue specific pattern that is required for UAS GFP expression to be driven.
Hugo Bellen
GFP inserts and Minos transformation.
BAC transformation up to 250kb, ‘no size limit’. Get 90% on first try.