Experiment: Inhibitory effect of Cas9 RNP alone on S10- or FSDS315-mediated GFP protein delivery to HeLa cells
PI: Paul B McCray, Jr, MD Alice F Tarantal, PhD David Guay, PhD David R Liu, PhD
Description: Inhibitory effect of Cas9 RNP alone on S10- or FSD315-mediated GFP protein delivery to HeLa cells. GFP protein (10 μM), S10 or FSDS315 (10 μM) with or without Cas9 RNP (containing 2.5 μM Cas9 and 2 μM gRNA) were added to HeLa cells and GFP delivery quantified by flow cytometry.
Delivery Assays:
Flow Cytometry
Parent Project: Base Editing in Rhesus Airway Epithelial
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Other experiments in this project: 6
- Delivery of Cas9 RNP using Shuttle peptide candidates to human airway epithelial cells cultured at the air liquid interface targeting CFTR locus
- Rhesus macaque CCR5 gRNA screening using ABE8e-Cas9
- Rhesus macaque CCR5 gRNA screening using ABE8e-Cas12a
- Delivery of ABE8e-Cas9 RNP using Shuttle peptide candidates to human airway epithelial cells cultured at the air liquid interface targeting B2M locus
- Delivery of ABE8e-Cas9 RNP using Shuttle peptide candidates to rhesus monkey airway epithelial cells cultured at the air liquid interface targeting CCR5 locus.
- Amphiphilic Peptides Deliver Base Editor RNPs to Rhesus Monkey Airway
Download: Submitted files
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Results |
| Delivery Efficiency | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Condition | Cell Type | Editor | Model | Delivery | Target Locus | Guide | Dosage | % of fluorescent cells | Image |
| Mock | HeLa | ||||||||
| GFP alone | HeLa | GFP protein (10μM) | |||||||
| S10 | HeLa | S10 | GFP protein (10μM) + Shuttle peptide (10μM) | ||||||
| S10 + Cas9 RNP | SpCas9 (Feldan Therapeutics) | HeLa | S10 | CFTR | Cas9-CFTR-R553X | GFP protein (10μM) + Shuttle peptide (10μM) + Cas9 RNP (2.5 μM Cas9 and 2 μM gRNA) | |||
| FSD315 | HeLa | FSD315 | GFP protein (10μM) + Shuttle peptide (10μM) | ||||||
| FSD315 + Cas9 RNP | SpCas9 (Feldan Therapeutics) | HeLa | FSD315 | CFTR | Cas9-CFTR-R553X | GFP protein (10μM) + Shuttle peptide (10μM) + Cas9 RNP (2.5 μM Cas9 and 2 μM gRNA) | |||
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