Experiment: Testing AAV5 for activation of tdTomato in mouse airway

PI: Guang-Ping Gao, PhD

Description: AAV2/5 mediated gene editing in the mouse airway was tested by deliverying SpCas9 and guide RNAs targeting the Ai9 transgene in Ai9 transgenic mice. Viral delivery was detected by GFP expression and gene editing quantified by tdTomato activation

Delivery Assays: 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 GFP. Delivery efficency was calculated by counting GFP+ cells compared to DAPI stained cells.

Editing Assay: 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.

Parent Project: Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases