Experiment: Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain
PI: Erik J Sontheimer, PhD
Description: Chemically modified crRNA and tracrRNA were injected into the striatum of TLR-MCV mice and the distribution of RNP was imaged.
Delivery Assays:
Delivery and uptake testing of C20-Cy3/T2-PS (red fluorescence) and SpyCas9 by immunohistochemical staining (green fluorescecnce) after intrastriatal injections
Parent Project: Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
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Other experiments in this project: 10
- Testing newly chemically modified crRNA and tracrRNA in mTmG mouse embryonic fibroblasts
- Testing newly chemically modified crRNA and tracrRNA in mouse Hepa 1-6 cells
- Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain
- Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
- Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
- Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
- Testing newly chemically modified crRNA and tracrRNA to activate the TLR reporter in human cells
- Testing newly chemically modified crRNA and tracrRNA to activate the TLR1 reporter in human cells
- Testing newly chemically modified crRNA and tracrRNA in mouse Neuro 2A cells
- Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center
Download: Submitted files
Results |
| Delivery Efficiency | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Condition | Tissue | Cell Type | Age | Editor | Model | Target Locus | Guide | Dosage | Injection Frequency | Signal | Image |
| Sp_c20_MCV1a/Sp_t2_5-3_PS-tail | brain | 7-8 weeks | SpyCas9-3xNLS | TLR-MCV1 mouse | TLR MCV reporter | STS118 (TLR-MCV1a; Sp_t2-PS:Sp_c20_TLR_MCV1a) | ~150 µM RNP | once |
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