Large Scale Projects
Generation of CRISPR resources to study phosphoinositide signaling in-vivo in a tissue specific manner
Fly Facility in collaboration with Raghu Padinjat, NCBS generated dual-gRNA (dgRNA) transgenic flies against all the genes known to interact with phosphoinositides. A set of >100 genes were selected and 2 gRNAs were designed, cloned, and tested in cell lines and flies for efficiency of deletion. These dgRNA transgenics were used for tissue specific genetic screens to study eye and wing development. More information on these can be obtained here
Establishment of a genome wide set of tagged fosmid reagents for the analysis of gene function in Drosophila melanogaster:
Fly Facility in collaboration with Pavel Tomancak, Frank Scnorrer, Mihail Sarov and other collaborators at MPI-CBG, Dresden and K. VijayRaghavan’s & Mani Ramaswami’s groups at NCBS generated transgenics for a large scale project “TransgeneOme”. We generated about 500 transgenic lines for the genomic clones of size ranging from 35-55kb by PhiC31 integrase mediated transgenesis.
The project aimed at expansion of the spectrum of available genomic resources by creating a comprehensive set of tagged proteins within their native genomic regulatory context. These reagents enable both the visualization of the tissue specificity and sub-cellular localization of proteins throughout the Drosophila life cycle and purification of protein-protein or protein-DNA complexes in a tissue specific manner.
More information on these can be obtained here
Drosophila protein interaction map (DPiM):
Fly Facility has generated about 5000 transgenic lines, for a large scale project DPiM, by injecting fly embryos with plasmid construct containing the ORFs fused to a HA tag sequence.
The goal of this project was to study how proteins encoded by Drosophila cDNAs interact with each other to form functional complexes. Out of the 13600 genes in the fly genome identified, ~5000 genes were fused to a HA tag to generate stable transgenic flies, from which the HA fused bait protein could be recovered along with its interacting partners, by Tandem Affinity Purification. The resulting protein complex could be studied through mass spectroscopic analysis, to identify the interacting partner. The expression of the bait protein in these flies is driven by a UAS promoter to allow spatio-temporal regulation. More information on these can be obtained here
DPiM fly stocks have now been discarded. However, the facility still maintains the plasmid DNA of these ORFs and can be obtained from the facility.
For more information on Fly (Drosophila) Facility, please write to services [at] ccamp.res.in or flyfacility [at] ccamp.res.in
Deepti Trivedi, In-Charge Fly Facility.