Experiment: Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center
PI: Erik J Sontheimer, PhD
Description: Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
  
  
  
  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, 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
- Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain
 
Download: Submitted files
  
    
| Results | 
| Editing Efficiency | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Condition | Tissue | Cell Type | Sex | Editor | Model | Target Locus | Guide | Injection Frequency | Signal | Image | 
| DNMT1-C20/T2 Control | striatum (TARGET) | Male | SpyCas9-3xNLS | mTmG mouse (congenic) | Dnmt1 | STS204 (DNMT1; Sp_t2:Sp_c20_Dnmt1) | once |  | ||
| mTmG-C20/T2 | striatum (TARGET) | Male | SpyCas9-3xNLS | mTmG mouse (congenic) | mTmG reporter transgene | STS159 (mTmG; Sp_t2:Sp_c20_mTmG) | once |  | ||
| Activation of mTmG reporter in mouse striatum | 
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| None provided. | 
