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PurposeStable 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.
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Depositing Labs
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Depositing OrganizationUniversity of Toronto
Located in Canada -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 221002 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 221002-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 221002-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonePB_tre_dCas9_VP160 (Addgene #126031)
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Vector typeMammalian Expression, CRISPR
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Selectable markersZeocin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namedCasRx-RBM25-IRES-BleoR
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Alt nameHuman RBM25 fused C-terminal of dCasRx, followed by IRES-BleoR selection cassette
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SpeciesH. sapiens (human)
- Promoter CAG
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Tags
/ Fusion Proteins
- NLS (N terminal on insert)
- NLS-HA (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
Resource Information
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A portion of this plasmid was derived from a plasmid made bydCasRx coding sequence from Konermann et al., 2018 (Addgene #109050)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Co-transfect with piggyBac transposase followed by Zeocin selection to generate polyclonal cells expressing dCasRx-RBM25. We recommend a high ratio of transposon to transposase for highest expression levels. Please see our paper for additional details.
DNA (Catalog # 221002-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 221002-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
PB-CAG-dCasRx-RBM25-IRES-BleoR was a gift from Benjamin Blencowe & Mikko Taipale (Addgene plasmid # 221002 ; http://n2t.net/addgene:221002 ; RRID:Addgene_221002) -
For your References section:
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. 10.1016/j.molcel.2024.05.028 PubMed 38917795
