psiRNA-DUO

Expression of Two shRNAs

psiRNA-DUO allows to generate two shRNAs from the same plasmid for the silencing of either a single target gene (with/without polymorphisms) or two different target genes. psiRNA-DUO contains special features designed to make the cloning of the shRNA inserts and selection of recombinant clone simple and rapid.

Specifications

Contains two 7SK RNAi cassettes

Allows white and blue selection

Features the GFP::zeo fusion gene

Provided with the psiRNA™ tools:
• 1 disk of lyophilized GT115 E.coli
• Sequencing primers (OL178-OL408 and OL906-OL176 pairs)
E. coli Fast-Media® XGal and E. coli Fast-Media® XGluc

Contents

psiRNA-DUO is provided as a kit composed of:

- 20 μg of psiRNA-DUO-GFPzeo
- 20 μg of psiRNA-LucLac-GFPzeo (control plasmid)
- 1 disk of lyophilized GT115 E. coli  (GT115 genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZM15 ΔlacX74 recA1 endA1 Δdcm uidA::pir-116 ΔsbcC-sbcD)
- 2 sets of sequencing primers (2 x 10 μg each)
- 2 pouches of Fast-Media® Zeo X-Gal
- 2 pouches of Fast-Media® Zeo X-Gluc

Description

The major features of the psiRNA-DUO plasmid are its two RNA Pol III cassettes and the GFP-Zeo fusion gene:

7SK RNAi Cassettes
The first cassette contains the following elements:
- Human 7SK promoter
- Acc65 I / Hind III cloning sites
- LacZ α-peptide expression cassette for white and blue selection

The second cassette contains the following elements:
- Human 7SK promoter
- Bbs I / Bbs I cloning sites
- GUS expression cassette for white and blue selection
β-Glucuronidase (GUS, uidA) is an E. coli enzyme capable of hydrolyzing the chromogenic substrate X-Gluc to generate a blue pigment. Therefore, similarly to LacZ, the expression of GUS in permissive E. coli allows the discrimination between blue parental clones and white recombinant clones.
psiRNA-DUO is provided with the E. coli GT115, a lacZ uidA mutant strain, enabling the white and blue color selection from the LacZ and GUS systems.

GFP-Zeo Fusion Gene
The GFP-Zeo gene is a transcriptional fusion between the Green Fluorescent Protein reporter gene and the Zeocin™ resistance (ZeoR) gene. The two moieties are separated by a bacterial promoter (EC2K) located within an intron. In E. coli, ZeoR is expressed from the EC2K promoter that is spliced out in mammalian cells. A composite CMV promoter drives the expression of the GFP-Zeo fusion gene, allowing the monitoring of transfection efficiency and the selection of stable clones.

Details

psiRNA-DUO backbone

Citations

Recent articles using InvivoGen psiRNA-DUO

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psiRNA-DUO

Cat. Codeksirna4-gz3
Unit Size1 kit
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Note that the sequence of available ORFs provided by InvivoGen can differ from a given reference Genbank record due to genetic variations and/or alternative splicing. Customers should verify that the version of a gene sold by InvivoGen is suitable for the customer needs.
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