Experiment: 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
PI: Erik J Sontheimer, PhD
Description: Chemically modified crRNA and tracrRNA were injected into the intra-striatum of mTmG reporter mice and activation of GFP expression was imaged.
Editing Assay:
Editing efficiency determined by immunohistochemical staining for GFP.
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, 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
- Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain
- Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center
Download: Submitted files
Results |
| Editing Efficiency | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Condition | Tissue | Cell Type | Age | Editor | Model | Delivery | Target Locus | Guide | Dosage | Injection Frequency | Signal | Image |
| Sp_c0_mTmG/Sp_t2 + S10 shuttle peptide | brain | 7-8 weeks | SpyCas9-3xNLS | mTmG mouse (congenic) | S10 | mTmG reporter transgene | STS159 (mTmG; Sp_t2:Sp_c0_mTmG) | 8 µM RNP + 40 µM shuttle peptide S10 | once |
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