Experiment Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice Experiment Condition ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9, female Assay Description: Editing efficiency is calculated by identifying airways in lungs sections by shape and DAPI-stained nuclei density. tdTomato-positive cells and total nuclei are counted in each airway. Airways are divided into small (<50000 square microns) and large (>50000 square microns) Tissue Measured: epithelium of main bronchus Cell Type : respiratory epithelial cell Measurement Type: Editing Efficiency Record ID 15000001078 Editor SpCas9 Delivery System Delivery System Subtype Guide BCM_ssAAV5-Sp_sgA; BCM_ssAAV5-Sp_sgB Vector ssAAV5-sgA+sgB-U1A.GFP; ssAAV5-spCas9; Model Species Mouse Model Name B6.Cg-Gt(ROSA)26Sor ^ tm9(CAG-tdTomato)Hze/J Application Method Intratracheal Intubation Application Site lung airway epithelium Dosage 1.7E11 viral genomes Injection Frequency once Injection Rate Injection Volume 40 microliters Days post injection 4 weeks Editor Format virus in sterile saline Antidote Id Antidote Description Measured Values Samples Size: 2   Replicate Result (Editing Efficiency) Measurement Units: % of cells Mean 19.95  1 19.3  2 20.6