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Experiment: Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter)

PI:   Aravind Asokan, PhD 

A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=5) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:4 ratio of cas9 to guide RNA (1e12vg of CMV Sacas9 vector and 3e12vg of the 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-Neg Control
Delivery Efficiency
Editing Efficiency
Target Tissue
Not Available

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