Cap-independent co-expression of dsRNA-sensing and NF-κB pathway inhibitors enables controllable self-amplifying RNA expression with reduced immunotoxicity
Lim TKY, Ritoux A, Paine LW, Ferguson L, Abdul T, Grundy LJ, Smith ESJ
bioRxiv 2024.09.24.614636
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Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 242407 | VEE-mScarlet3-IRES-EGFP-IRES-moxBFP | Native (conventional) self-amplifying RNA (saRNA) control construct expresses moxBFP as visual marker of IRES functionality |
| 242408 | VEE-mScarlet3-IRES-EGFP-IRES-E3 | Self-amplifying RNA (saRNA) E3 construct; expresses the vaccinia virus E3 protein, which binds and sequesters dsRNA broadly inhibiting dsRNA-sensing pathways |
| 242409 | VEE-mScarlet3-IRES-EGFP-IRES-E3-NSs-L* | Self-amplifying RNA (saRNA) E3-NSs-L* construct; co-expresses vaccinia virus E3, Toscana virus NSs, and Theiler’s virus L*; more comprehensive inhibition of dsRNA-sensing pathways than plasmid 242408. |
| 242410 | VEE-mScarlet3-IRES-E3-NSs-L*-EGFP-IRES-moxBFP | Self-amplifying RNA (saRNA) construct expresses dsRNA pathway inhibitors (vaccinia virus E3, Toscana virus NSs, Theiler’s virus L*) and moxBFP (in place of inflammatory signaling inhibitors). |
| 242411 | VEE-mScarlet3-IRES-E3-NSs-L*-EGFP-IRES-srIκBα | Self-amplifying RNA (saRNA) construct expresses dsRNA pathway inhibitors (E3, NSs, L*) and inflammatory signaling inhibitor, super repressor inhibitor of κBα (srIκBα). |
| 242412 | VEE-mScarlet3-IRES-E3-NSs-L*-EGFP-IRES-srIκBα-Smad7-SOCS1 | Self-amplifying RNA (saRNA) construct expresses dsRNA pathway inhibitors (E3, NSs, L*) and inflammatory signaling inhibitors, srIκBα, Smad7, and SOCS1. |
| 242413 | VEE-moxBFP-IRES-PuroR | Spectral unmixing: moxBFP only |
| 242414 | VEE-EGFP-IRES-PuroR | Spectral unmixing: EGFP only |
| 242415 | VEE-mScarlet3-IRES-PuroR | Spectral unmixing: mScarlet3 only |