Experiment Testing AAV5 for activation of tdTomato in mouse airway club and ciliated cells Experiment Condition Dual AAV Assay Description: Editing efficiency calculated by counting cells double positive for tdTomato and CCSP (a marker of club/clara cells) and acetylated tubulin (a marker of ciliated cells) compared to DAPI stained cells. Analyzed 5 sections per mouse. Tissue Measured: epithelium of bronchus Cell Type : club cell Measurement Type: Editing Efficiency Record ID 15000001640 Editor SpCas9 Delivery System Delivery System Subtype Guide sgAi9L; sgAi9R Vector AAV.pU1a-SpCas9; H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1); Model Species Mouse Model Name B6.Cg-Gt(ROSA)26Sor ^ tm9(CAG-tdTomato)Hze/J Sample Age 8 weeks Sample Sex Female Application Method inhalation Application Site airway epithelium Dosage 6e10 gc/each AAV per mouse by intratracheal injection Injection Frequency once Injection Rate n/a Injection Volume 40 microliters Days post injection 4 weeks Editor Format AAV vector encoded Antidote Id Antidote Description Antibody RRID Antibody Other ID Thermo Fisher Scientific, MA5-15257 Antibody Description Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300 Antibody RRID Antibody Other ID Santa Cruz, SC-25555 Antibody Description Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000 Antibody RRID Antibody Other ID Santa Cruz, SC-23950 Antibody Description Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200 Antibody RRID Antibody Other ID Rockland, 600- 401-379 Antibody Description Anti-RFP (Rabbit) Polyclonal Antibody Measured Values Samples Size: 3   Replicate Result (Editing Efficiency) Measurement Units: % cells Mean 14.97  1 15.2  2 12.5  3 17.2