EF1a-dCasRx-RBM25
(Plasmid #221001)

• Purpose: Transient expression of dCasRx-RBM25. EF1a promoter.
• Depositing Labs: Benjamin Blencowe, Mikko Taipale
• Publication: Efficient, specific, and combinatorial control of endogenous exon splicing with dCasRx-RBM25 (unpublished)

Backbone

• Vector backbone: pXR002 (Addgene #109050)
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: dCasRx-RBM25
• Alt name: Human RBM25 fused C-terminal of dCasRx
• Species: H. sapiens (human)
• Promoter: EF1a
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • NLS-HA (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Backbone and dCasRx coding sequence from Konermann et al., 2018 (Addgene #109050)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Co-transfect with gRNA vector at a 1:1 ratio. Highly efficient transfection is required for optimal transient splicing modulation. For hard to transfect cells we recommend either sorting for BFP expression (on the gRNA vector), or using the stable expression system. For additional details, please see our paper.