pAAVS1-SA-T2A-Neo-CAG-rtTA-pA-TRE-dCas9-KRAB-pA Donor Vector
| Catalog# | Unit | Unit Price (USD) | Actions |
|---|---|---|---|
| AI7902 | 10 μg at 0.5 μg/μL in TE | $1,250.00 | Add to Cart |
Description:
CRISPR interference (CRISPRi) stands out as a powerful tool for targeted gene transcription silencing in both prokaryotic and eukaryotic cells. The collaboration between inactive Cas9 (dCas9) and the Krüppel-associated box (KRAB) repressor ensures accurate and reversible gene silencing. Additionally, the strategic use of the Human AAVS1 "safe harbor" site, located within intron 1 of the PPP1R12C gene, minimizes its impact on cellular functions. Crucially, our approach incorporates the renowned TeTOn gene inducible system, providing precise control over gene expression. Responsive to specific signals, such as tetracycline, this system allows researchers to activate or deactivate genes as needed. Our innovative pAAVS1-SA-T2A-Neo-CAG-rtTA-pA-TRE-dCas9-KRAB-pA donor vector seamlessly integrates the strengths of CRISPRi, the AAVS1 safe harbor, and the TetON gene inducible system. Featuring a TRE promoter-controlled CRISPRi-KRAB fusion gene, this design ensures manipulable and robust expression of the dCAS9/KRAB protein with or without tetracycline treatment, without disrupting cellular functions. The vector strategically integrates the Neomycin selection marker with a splice acceptor (SA) site within the AAVS1 safe harbor locus, closely linking Neomycin expression to intron integration. This significantly reduces the risk of unintended off-target integrations during Neomycin selection. When combined with the pSpCAS9 nuclease & AAVS1 gRNA vector, the pAAVS1-SA-T2A-Neo-CAG-rtTApA-TRE-dCas9-KRAB-pA donor vector facilitates the seamless integration of a TeTOn-controlled dCAS9/KRAB fusion protein into the AAVS1 safe harbor site. This approach, along with specific gene-targeted gRNAs, enables inducible and precise and highly efficient repression of the target gene expression, providing an accessible and consistent method for specific gene silencing.
Vector Information
This AAVS1 donor vector includes essential components to serve as a reliable integration marker in the AAVS1 Safe Harbor site. It incorporates a universal SA-T2A-NeoR cassette, facilitating the integration of a dCas9/KRAB fusion gene driven by an inducible TRE promoter into the AAVS1 safe harbor site through antibiotic selection. This vector is suitable for generating inducible CRISPRi stable cell lines.
IMPORTANT NOTICE
Store the vial at -20°C immediately upon receipt.
Specifications:
| Product Name | pAAVS1-SA-T2A-Neo-CAG-rtTA-pA-TRE-dCas9-KRAB-pA Donor Vector |
|---|---|
| Shipping Condition |
Room Temperature - 2 Day Shipping |
| Storage and Stability |
Store at -20°C immediately upon receipt. This product is stable for 6 months when stored as directed. |
| Quality Control | This plasmid is sequence verified. |
| Restricted Use |
For Research Use Only. Not for use in diagnostic or therapeutic procedures. |