pAAVdual-CAG-iCre

Price range: $999.00 through $1,899.00

pAAVdual-CAG-iCre  is used to produce AAV-CAG-iCre virus using AAVnerGene’s novel AAVdual production system. In this plasmid,  the  regular single strand AAV genome  with a CAG promoter and an iCre reporter is cloned into our novel Ad helper plasmid, mini-pHelper.  AAV-CAG-iCre viruses can be generated by co-transfection of this plasmid with regular AAV helper  (pRCap) plasmids, carrying AAV2 rep gene and different cap genes,  without adding additional Ad helper plasmid to supply E2A, E4orf6 and VA RNA functions.

The CAG promoter is a synthetic hybrid promoter widely used in molecular biology to drive high levels of gene expression in mammalian cells. It is composed of three key elements:CMV Immediate-Early Enhancer: Derived from the cytomegalovirus (CMV), this enhancer sequence provides strong and ubiquitous transcriptional activation in a variety of cell types. Chick Beta-Actin Promoter: The core promoter region is derived from the chicken beta-actin gene, which is known for its high expression levels in many cell types, particularly in the context of gene expression in vertebrates. Rabbit Beta-Globin Intron: An intronic sequence from the rabbit beta-globin gene is included to enhance transcriptional efficiency and mRNA stability, further boosting expression levels. It is particularly valued for its strength and broad activity across various cell types, making it a popular choice for applications such as gene therapy, transgenic animal models, and research studies that require robust and consistent gene expression.

iCre is a codon-optimized variant of Cre recombinase, designed to enhance its expression and efficiency in mammalian cells. Like the standard Cre recombinase, iCre recognizes loxP sites and catalyzes site-specific recombination between them, which can result in the deletion, inversion, or translocation of the DNA sequence flanked by these sites. AAV-iCre offers an improved and more efficient approach to conditional gene manipulation, providing researchers with a powerful tool for studying gene function, lineage tracing, and gene editing. 

  • AAV Cassette: AAV-CAG-iCre
  • AAV Genome Size: 2854 bp
  • Plasmid Size: 11287 bp
  • Plasmid Backbone: pAAVdual
  • AAV Genome Type: ssAAV
  • ITRs:  L-ITR (119 bp) + R-ITR (130 bp)
  • Promoter: CAG
  • Transgene: iCre
  • Bacterial Resistance(s) : Kana
  • Storage:  -20°C for long term (> 1 year)
  • Shipping: Dry ice.
  • Notice to Purchaser:  MTA is required. All mini-pHelper based products, including mini-pHelper itself, pAAVdual and pAAVone plasmids,  are sold under license of US 20240229067. Rights to use these products are limited to non-commercial research only. No other rights are conveyed. Inquiry into the availability of a license to broader rights or the use of these products for commercial purposes should be directed to AAVnerGene Inc, 9620 Medical Center Dr, Suite 100, Rockville, MD, 20850. Purchase of these products does not grant rights to : (1) offer the vectors or any derivatives thereof for resale; or (2) to distribute or transfer the vector or any derivative thereof to third parties.
  • 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 (CsCl, Iodixanol, or AAV-X resin).
  • AAV Analytical Services: We also offer advanced AAV characterization services,  including AAV purity, vector genome identity, ITR integrity, DNA impurities, empty/full ratios, and a variety of comprehensive analyses based on custom requests.
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pAAVdual plasmids are used to produce regular single strand AAV (ssAAV) viruses using AAVnerGene’s novel AAVdual System. In this series plasmids, the AAV genomes are cloned into our novel Ad helper plasmid, mini-pHelper.  AAV viruses can be generated by co-transfection of pAAVdual plasmids  with the regular AAV helper (pRCap) plasmids carrying different AAV serotypes.

AAV-iCre Viruses with Different Serotypes and Promoters: 

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pAAVdual-Pro-iCre Plasmids:

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AAVdual System vs Traditional Triple-Plasmid System:

  • Achieve up to a 250% increase in AAV yield.
  • Reduce the plasmid count from 3 to 2.
  • Simplify plasmid ratio optimization and minimize batch-to-batch variations.
  • Allow flexibility in packaging AAVs with different capsids.
  • Utilize a compact plasmid size (8.5 kb plasmid backbone + AAV genome) with comparable plasmid yields.
  • Maintain or improve the full-to-empty capsid ratio.
  • Ensure similar viral activity, capsid composition, and genomic integrity.
  • Maintain consistent levels of rcAAV, hcDNAs, and producer plasmid contamination.
  • Preserve the same upstream and downstream processes, including analytical release and characterization assays.
This plasmid  offers a relative flexible, simple, low cost, but high consistency and high efficiency for production of AAV virus with different AAV serotypes.

 

AAVone, AAVdual, AAVtri and traditonal triple-plasmid AAV production system
AAVone, AAVdual, AAVtri and traditional triple-plasmid AAV production systems
Amount

10 ug, 100 ug, 1 mg, 10 mg