Efficient, specific, and combinatorial control of endogenous exon splicing with dCasRx-RBM25.
Li JD, Taipale M, Blencowe BJ
Mol Cell. 2024 Jul 11;84(13):2573-2589.e5. doi: 10.1016/j.molcel.2024.05.028. Epub 2024 Jun 24.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 221001 | EF1a-dCasRx-RBM25 | Transient expression of dCasRx-RBM25. EF1a promoter. |
| 221002 | PB-CAG-dCasRx-RBM25-IRES-BleoR | Stable expression of dCasRx-RBM25 when co-transfected with piggyBac transposase. Zeocin selection. CAG promoter. Use with lentiviral gRNA delivery (Addgene #138150) to stably modulate splicing. |
| 221003 | CMV-dCasRx-ccdB | Destination vector for Gateway cloning of ORFs into the C-terminus of dCasRx. CMV promoter. Transient expression. |
| 221004 | gRNA-CasRx_SD1-pSV40-TagBFP | Transiently express CasRx DR30 repeat with gRNA spacer. Contains SD1 mutation to enhance efficiency. SV40-TagBFP cassette to monitor transfection efficiency. Use BbsI with overhangs Fw-AAAC, Rv-AAAA. |
| 221005 | SMN2E7-Fluc-Nluc-reporter | Transiently express SMN2 dual luciferase reporter. Inclusion of E7 produces in-frame stop codon and Firefly luciferase only, while skipping produces both Firefly and Nanoluc. CMV promoter. |