The UAS-cDNA/ORF constructs generated by CRISPRmass were expressed in the presence of GAL4 in Drosophila. Transgenic UAS-CG7747, UAS-Prdm13, and 38 UAS-DUB lines were generated by CRISPRmass in Drosophila using the 40 cDNA/ORF plasmids constructed in eight distinct vectors. An epitope-tagged transgene for each of the eight vectors was chosen and crossed to motor neuron–specific OK6-GAL4 and eye-specific GMR-GAL4 drivers, respectively. (DUB) Deubiquitinase. (A,B) Immunostaining showed these transgenes were expressed in the larval motor neurons, and V5 signals are indicated by arrows. Presynaptic active zones are labeled with anti-BRP (nc82) (red). (C–E) Western blotting showed these transgenes were expressed in adult fly eyes. The epitope-tagged proteins and their expected molecular weights are as follows: CG4968-3MYC, 33.9 kD; Rpn8-3MYC, 41.6 kD; Usp5-3MYC, 95.6 kD; CG7747-3MYC, 62.5 kD; Usp20-33-3MYC, 113.2 kD; Prdm13-3MYC, 54.1 kD; Ctbp2-2MYC, 48.3 kD; USP8-V5, 130.0 kD.
