Cat. #161705
Dicer activation reporter plasmid (DARA plasmid)
Cat. #: 161705
Bacterial Resistance: Ampicillin
Selectable Markers: Hygromycin
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Contributor
Inventor: Andrii Domanskyi, Piotr Chmielarz, Ilmari Parkkinen
Institute: University of Helsinki
Primary Citation: N/A
Tool Details
*FOR RESEARCH USE ONLY
- Tool name: Dicer activation reporter plasmid (DARA plasmid)
- Alternate name: pTO-miR19b-tdTom-19b_bind-WPRE-EGFP Dicer-activating reporter assay (DARA) plasmid
- Purpose: Plasmid for creation of DARA reporter cell line, by homologous recombination in HEK293 Flp-In T-REx cells
- Disease: Parkinson's disease; ALS
- Insert: Fluorescent proteins tdTomato and EGFP; human miR-19a-1 hairpin and binding site
- Total vector size: 9990 bp
- Backbone size without insert: 4329 bp
- Insert size: 5661 bp
- Insert species: Aequorea victoria; Homo sapiens
- Promoter: Tet-ON CMV; SV40; TK-HSV
- Bacterial resistance: Ampicillin
- Selectable markers: Hygromycin
- Description: DARA plasmid was created for the Dicer activation reporter assay, which is used to screen compounds for miRNA biogenesis activation. Dicer has been implicated in certain cancers, ALS and Parkinsons Disease. The DARA plasmid was designed to express three main elements: 1) a primary fluorescent reporter protein, a tandem dimer Tomato (tdTomato) red fluorescent protein, which additionally has a nuclear localization sequence (NLS), 3’ untranslated sequence (3’UTR) containing miRNA miR-19b bin...
- Application: High throughput screening of chemical compound libraries
- Cloning information: miRNA-19b-1 hairpin (184 bp) was amplified from human genomic DNA with the folowing primers and cloned into psiCHECK vector (digested with SalI/ApaI) using inFusion system.
miR19b_for: 5’ gaggtagtgagtcgacGCCCAATCAAACTGTCCTGTTACTG
miR19b_rev: 5’ catcagcgaaccgcgggcccGGTTTGAGTTTCCCTTACTTTTCTACAGAC
Handling
- Storage conditions: Store at -20° C
- Storage buffer: Water
- Shipping conditions: +4° C or dry ice
- Plasmid amplification details: Stbl3 E.coli
- Plasmid copy number: High copy number
Application Details
- Application: High throughput screening of chemical compound libraries
Related Tools
- Related tools: Dicer activation reporter cell line (161704)
References
- N/A
