pAAVtri-H1-shRNA-EF1a-EGFP

pAAVtri-H1-shRNA-EF1a-EGFP plasmid is designed to produce AAV-shRNA vectors, in which the shRNA is  under the control of the U6 Pol III promoter. Users can clone their shRNA into this plasmid by replacing the shRNA-scramble cassette between the HindIII and MluI sites. The EF1a-EGFP expression cassette, located downstream of the H1-shRNA cassette, allows for monitoring transfection efficiency. To generate AAV-shRNAs (AAV-H1-shRNA-EF1a-EGFP) with the desired AAV serotypes, users can co-transfect this plasmid with the pRCap plasmid (which carries the AAV rep and cap genes) and the mini-pHelper-1.0 plasmid (which supplies adenoviral E2A, E4orf6, and VA RNA functions) into HEK 293T cells, using our AAVtri AAV production system.

• Full Plasmid Name and Sequence: pAAVtri-H1-shRNA-Scramble-EF1a-SV40I-EGFP-WPRE3-SV40polyA
• Plasmid Size: 5415 bp
• Plasmid Backbone: pAAVtri
• AAV Genome Type: ssAAV
• ITRs: L-ITR (119 bp) + R-ITR (130 bp)
• shRNA Promoter: H1
• shRNA-scramble: 5‘-CCTAAGGTTAAGTCGCCCTCG-3’
• Reporter Expression Cassette: EF1a-EGFP
• Bacterial Resistance(s) : Kana
• Storage:  -20°C for long term (> 1 year)
• Shipping: Ice bag
• Cloning Services: AAVnerGene provides cloning services based on this plasmid backbone with $300/clone.
• AAV Package Services: AAVnerGene provides affordable AAV production services at various scales(from 1e12 to 1e16 vg), in both suspension and adherent cultured cells (HEK 293T or HEK 293), with flexible purification methods (Iodixanol, CsCl, or AAV-X resin).
• AAV Analytical Services: We also offer advanced AAV characterization services,  including Full/empty ratios, AAV purity, AAV potency, vector genome identity, ITR integrity, DNA impurities, rcAAV, and a variety of comprehensive analyses based on custom requests.