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Experiment: Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CMV promoter) and self complementary sgRNA vector.

PI:   Aravind Asokan, PhD 
 

A dual vector strategy was employed: one self complementary vector delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (single stranded vector). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=4) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:1 ratio of cas9 to guide RNA (2e12vg of CMV Sacas9 vector and 2e12vg of the self complementary sgRNA vector) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.
Editing Assay: - Editing efficiency calculated by counting fluorescent cells (TdTomato) compared to DAPI stained cells.

Organ System Overview


 
AAV9, Male
AAV9, Female
AAVcc47, Male
AAVcc47, Female
Mock
Legend:  
Delivery Efficiency
Editing Efficiency
Target Tissue
Not Available
 

 
Analyze Data Sets Available for this Experiment
Liver and Biliary
  Liver (unspecified)
Cardiovascular
  Heart (unspecified)