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ExperimentTesting AAV5 for activation of tdTomato in mouse airway club and ciliated cells
Experiment ConditionAAV-SpCas9
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 ID15000001638

EditorSpCas9
Delivery System
Delivery System Subtype
Guide
VectorAAV.pU1a-SpCas9;

Model SpeciesMouse
Model NameB6.Cg-Gt(ROSA)26Sor ^ tm9(CAG-tdTomato)Hze/J

Application Methodinhalation
Application Siteairway epithelium
Dosage6e10 gc/each AAV per mouse by intratracheal injection
Injection Frequencyonce
Injection Raten/a
Injection Volume40 microliters
Days post injection4 weeks
Editor FormatAAV vector encoded
Antidote Id
Antidote Description

Antibody RRID
Antibody Other IDThermo Fisher Scientific, MA5-15257
Antibody DescriptionAnti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300

Antibody RRID
Antibody Other IDSanta Cruz, SC-25555
Antibody DescriptionAnti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000

Antibody RRID
Antibody Other IDSanta Cruz, SC-23950
Antibody DescriptionAnti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200

Antibody RRID
Antibody Other IDRockland, 600- 401-379
Antibody DescriptionAnti-RFP (Rabbit) Polyclonal Antibody

Measured Values
Samples Size: 3
 
Replicate Result (Editing Efficiency)
Measurement Units: % cells
Mean 0.0 
1
2
3
  

Publication Title
Cross-species evolution of a highly potent AAV variant for therapeutic gene transfer and genome editing. NCBI