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Benjamin Blencowe Lab: Efficient, specific, and combinatorial control of endogenous exon splicing with dCasRx-RBM25

Li J.D., Taipale M., Blencowe B.J.
Mol Cell, 2024 (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
221001EF1a-dCasRx-RBM25Transient expression of dCasRx-RBM25. EF1a promoter.
221002PB-CAG-dCasRx-RBM25-IRES-BleoRStable 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.
221003CMV-dCasRx-ccdBDestination vector for Gateway cloning of ORFs into the C-terminus of dCasRx. CMV promoter. Transient expression.
221004gRNA-CasRx_SD1-pSV40-TagBFPTransiently 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.
221005SMN2E7-Fluc-Nluc-reporterTransiently 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.

Antibodies from Article