21nt siRNA oligos were designed to target exons common to all isoforms of a gene using the DSIR website tool. Oligos were compared to SNP resequencing data from 10 Drosophila genomes. For each gene, the best siRNA oligo was selected on a combination of highest predicted activity, lowest off-target probability and absence of common SNPs in a critical position. Any shRNAs exactly matching those at the DRSC TRiP resource were eliminated. Each of the siRNA oligos was then incorporated into a 71nt oligo to generate shRNA (short RNA hairpin). This was cloned pWALIUM20 vector to enable GAL4-inducible expression in germline and somatic cells. Constructs were integrated into the attP40 genomic landing site on the left arm of the second chromosome at 25C6, which has been shown to provide high levels of induced expression of transgenes, yet maintain a low basal expression when the transgenes are not induced.

21nt siRNA oligos were designed to target exons common to all isoforms of a gene using the DSIR website tool. Oligos were compared to SNP resequencing data from 10 Drosophila genomes. For each gene, the best siRNA oligo was selected on a combination of highest predicted activity, lowest off-target probability and absence of common SNPs in a critical position. Any shRNAs exactly matching those at the DRSC TRiP resource were eliminated. Each of the siRNA oligos was then incorporated into a 71bp oligonucleotide to generate shRNA (short RNA hairpin) as follows. A top strand oligo was designed by concatenating "ctagcagt", the sense-strand oligo (21 nucleotides), "tagttatattcaagcata", the anti-sense oligo, and "gcg". A bottom strand oligo was designed by concatenating "aattcgc", the sense-strand oligo,"tatgcttgaatataacta", the anti-sense oligo, and "actg". This 71bp oligonucleotide was cloned pWALIUM20 vector to enable GAL4-inducible expression in germline and somatic cells. Constructs were integrated into the attP40 genomic landing site on the left arm of the second chromosome at 25C6, which has been shown to provide high levels of induced expression of transgenes, yet maintain a low basal expression when the transgenes are not induced.