A cassette comprised of the Tag:V5 epitope plus the Tag:miGFPi RNAi-effector sequence has been inserted in frame into the common N-terminal domain of lola. The position of the cassette in the common N-terminal sequence allows for conditional knockdown of all lola isoforms using an RNAi reagent that targets EGFP.
Expression of Avic\GFPdsRNA.shRNA.Scer\UAS.1 under the control of Scer\GAL4wor.PA in lolaV5.miGFPi.all homozygotes to knock-down all isoforms of lola using the miGFPi method (tagging a gene with a minimal GFP tag and then expressing RNAi against the tag) results in overproliferation of neural stem cells (at the expense of differentiating cells) in the larval central brain. Avic\GFPdsRNA.shRNA.Scer\UAS.1 expression in lolaV5.miGFPi.all/lolaV5.miGFPi.PD (effectively knocking down lola-PD only) does not cause overproliferation of neural stem cells in the larval central brain, but when both lola-PD and lola-PB isoforms are knocked-down simultaneously in lolaV5.miGFPi.all/lolaV5.miGFPi.PD,lolaHA.miGFPi.PD larvae, neuroblast overproliferation is observed.