Experiment: Testing new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in primary fibroblast cells from adult Ai14 mouse cochlea
PI: Zheng-Yi Chen, PhD
Description: LNP delivery of Cas9 mRNA/sgRNA in primary fibroblast cells from adult Ai14 mouse cochlea (inner ear)
Editing Assay:
Editing is displayed by tdTomato signal 48 hr after transfection.
Parent Project: Efficient In Vivo RNP-Based Gene Editing in the Sensory Organ Inner Ear Using Bioreducible Lipid Nanoparticles
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Other experiments in this project: 3
- Verification of new LNP 306-O12B for delivery of Cas9 mRNA/sgRNA in adult mouse cochlea. Procedure performed side by side with JAX.
- Testing new LNPs (lipid nanoparticles) for delivery of Fluc mRNA in adult mice
- Testing new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in adult mouse cochlea
Download: Submitted files
Results |
| Editing Efficiency | ||||||||
|---|---|---|---|---|---|---|---|---|
| Condition | Editor | Model | Delivery | Target Locus | Guide | Dosage | Signal | Image |
| 306-O12B | CleanCap® Cas9 | Ai14 mouse primary fibroblasts | 306-O12B | Ai9/Ai14 reporter transgene | sg298 | LNP/Cas9/sgRNA=15/1/1, [LNP]=7.6ug/ml | ||
| 306-S10 | CleanCap® Cas9 | Ai14 mouse primary fibroblasts | 306-S10 | Ai9/Ai14 reporter transgene | sg298 | LNP/Cas9/sgRNA=15/1/1, [LNP]=7.6ug/ml | ||
| LNP delivery of Cas9 mRNA/sgRNA in primary fibroblast cells from adult Ai14 mouse cochlea (inner ear) |
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In vitro screening of LNPs. Primary fibroblast cells were derived from skins of P5 Ai14 mice and plated in 48-well plates at 10,000-20,000 cells per well. LNP was mixed with Cas9 mRNA and sgRNA in 15:1:1 ratio and added to the well. The final concentration of LNP is 7.6 ug/ml. Cells were collected 48 hr later to check tdT signal. a, 306-O12B; b, 306-S10 |
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