Experiment: Podocyte-specific gene editing in human kidney organoids
PI: Benjamin S Freedman, PhD Ross C. Wilson, PhD
Description: Kidney organoids were derived from a human iPS cell line with Ai9 (tdTomato) fluorescence-on reporter knocked into the AAVS1 safe harbor locus. Intact kidney organoids were transfected with CRISPR ribonucleoprotein complexes with and without molecular targeting agent (MTA) specific for podocytes. Genome editing events were detected by induction of tdTomato from the Ai9 reporter.
Editing Assay:
Calculated Editing efficiency by counting fluorescent cells (TdTomato) compared to Wilms Tumor-1 (WT1) stained cells. NOTE: Each data point represents one organoid.
Parent Project: Cas9 ribonucleoprotein delivery targeted to kidney epithelium
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Other experiments in this project: 1
Download: Submitted files
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Results |
| Editing Efficiency | ||||||||
|---|---|---|---|---|---|---|---|---|
| Condition | Sex | Editor | Model | Delivery | Target Locus | Guide | WT1+tdTomato+/total tdTomato+ (%) | Image |
| RNP | Female | SpyCas9-ProteinG-3xNLS | human kidney organoid with Ai9 (tdTomato) fluorescence-on reporter | endosomolytic peptide | Ai9/Ai14 reporter transgene | sg298 | ||
| RNP + MTA | Female | SpyCas9-ProteinG-3xNLS | human kidney organoid with Ai9 (tdTomato) fluorescence-on reporter | endosomolytic peptide and anti-podocyte targeting antibody | Ai9/Ai14 reporter transgene | sg298 | ||
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