Experiment: Testing newly chemically modified crRNA and tracrRNA to activate the TLR reporter in human cells
PI: Erik J Sontheimer, PhD| Chemically modified crRNA and tracrRNA were delivered by electroporation in presence of amphiphilic peptide to transgenic human HEK-293T cells harboring the TLR-MCV1 reporter. Gene editing was determined by reporter activation. |
| Editing Assay: |
- Calculated editing efficiency by flow cytometry analysis of RFP expression. |
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Results |
| Editing Efficiency | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Condition | Sex | Editor | Model | Delivery | Target Locus | Guide | Dosage | % of RFP+ cells | Image |
| STS118 (TLR-MCV1a) (5 pmol RNP) | Female | SpyCas9-3xNLS | HEK-293T-disrupted_GFP with MCV-mcherry-Puro | FSD117D1 | TLR MCV reporter | STS118 (TLR-MCV1a; Sp_t2:Sp_c20_TLR_MCV1a) | 5 pmol RNP | ||
| STS118 (TLR-MCV1a) (20 pmol RNP) | Female | SpyCas9-3xNLS | HEK-293T-disrupted_GFP with MCV-mcherry-Puro | FSD117D1 | TLR MCV reporter | STS118 (TLR-MCV1a; Sp_t2:Sp_c20_TLR_MCV1a) | 20 pmol RNP | ||
| STS119 (TLR-MCV1b) (20 pmol RNP) | Female | SpyCas9-3xNLS | HEK-293T-disrupted_GFP with MCV-mcherry-Puro | TLR MCV reporter | STS119 (TLR-MCV1b; Sp_t2:Sp_c20_TLR_MCV1b) | 20 pmol RNP | |||
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