A cassette comprised of the Tag:V5 epitope plus the Tag:miGFPi RNAi-effector sequence has been inserted at the N-terminal end of the Tif-IA coding sequence. The presence of the Tag:miGFPi tag allows for conditional knockdown of Tif-IA function using an RNAi reagent that targets EGFP.
Expression of Avic\GFPdsRNA.shRNA.Scer\UAS.1 RNAi under the control of Scer\GAL4nos.UTR.T:Hsim\VP16 in Tif-IAV5.miGFPi homozygotes to knock-down Tif-IA by the miGFPi method (tagging a gene with a minimal GFP tag and then expressing RNAi against the tag) results in a complete loss of germline stem cells (and consequently all germline cells) and empty ovarioles in adult females.
Tif-IAV5.miGFPi is partially rescued by Tif-IAS610A.UASp/Scer\GAL4VP16.nanos.UTR/Avic\GFPRNAi.UAS.V20.1
The loss of germline stem cells phenotype observed in ovarioles of adult Tif-IAV5.miGFPi females upon Scer\GAL4nos.UTR.T:Hsim\VP16-driven knock-down of Tif-IA using the miGFPi method (RNAi is targeted against the GFP tag inserted in a particular gene locus) is fully rescued by co-expression of Tif-IAScer\UAS.P\T.cWa.