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Cas13bt3 knockdown of VEGFA mRNA in mammalian cells and mouse retina [RNA-Seq]

GSE259220 Homo sapiens; Mus musculus Expression profiling by high throughput sequencing 61 samples 2024/03/01 GPL24676GPL24247
Summary
Purpose: We report expression of a new Cas13 enzyme in retinal cells and evaluate its efficacy in targeting the VEGFA mRNA to establish safety and efficacy of the enzyme for anti-VEGFA therapy. Methods: Mammalian cells (HEK293FT) or mouse retina were treated with shRNA, CasRx or Cas13bt3 carrying VEGFA targeting sgRNA. Total RNA was collected for RNA seqeuencing to study VEGFA mRNA expression and well as off-target genes. Results: Significant knockdown of VEGFA mRNA was observed with no effect on predicted off-target genes in mammalina cells and mouse retina. No loss of mouse VEGFA was also detected. Conclusions: AAV2.7m8 can be used for retina-wide transduction, and Cas13bt3 may be a potential new tool for control of VEGFA.
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