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ExperimentTesting AAV5 for activation of tdTomato in mouse airway
Experiment ConditionDual AAV
Assay Description: Two AAVs encoding sgRNAs and SpCas9 were introduced by intratracheal injection into Ai9 mice. After 4 weeks, lung tissue was harvested and prepared by sectioning and immunohistochemical staining for tdTomato. Editing efficiency calculated by counting fluorescent cells (RFP) compared to DAPI stained cells.
Tissue Measured:epithelium of main bronchus
Cell Type :epithelial cell of lung
Measurement Type:Editing Efficiency
Record ID15000001636

EditorSpCas9
Delivery System
Delivery System Subtype
GuidesgAi9R; sgAi9L
VectorAAV.pU1a-SpCas9; H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1);

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 IDCell Signaling Technology Cat# 2956
Antibody DescriptionGFP (D5.1) XP Rabbit mAb antibody

Antibody RRID
Antibody Other IDRockland Cat# 600-401-379
Antibody DescriptionAnti-RFP (RABBIT) Antibody

Measured Values
Samples Size: 5
 
Replicate Result (Editing Efficiency)
Measurement Units: % cells
Mean 24.2 
1 20.5 
2 21.5 
3 25.4 
4 23.1 
5 30.5 
  

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