Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice |
ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, male |
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) |
epithelium of main bronchus |
respiratory epithelial cell |
Editing Efficiency |
15000001049 |
|
SpCas9 |
|
|
BCM_ssAAV5-Sp_sgA |
ssAAV5-sgA+sgB-U1A.GFP; ssAAV5-spCas9; |
|
Mouse |
B6.Cg-Gt(ROSA)26Sor^em1(CAG-tdTomato)JaHe |
|
Intratracheal Intubation |
lung airway epithelium |
1.7E11 viral genomes |
once |
|
40 microliters |
4 weeks |
virus in sterile saline |
|
|
|
Measured Values |
Samples Size: | 1 |
|
Mean |
17.6  |
1 |
17.6  |