The ATHENA I AAV Capsid Platform is a recognized capsid library employed to methodically assess AAV serotypes or variations for diverse objectives with Barcode-seq technology. This platform comprises a repository of more than 1000 distinct AAV capsids, with each capsid variant being linked to three unique DNA barcodes. Through the comparative analysis of DNA or RNA levels associated with these barcodes, researchers can ascertain the AAV capsid variant that demonstrates the highest efficiency tailored to their particular application.
Purposes of ATHENA I
AAV vectors represent potent tools in gene delivery and gene therapy. With the discovery of numerous natural AAV serotypes and the growing number of engineered variants, the quest for superior capsids tailored to specific cell types and tissues is crucial for effective gene therapies. However, a practical method for achieving this has been lacking. Presently, the typical approach involves individually testing AAV variants, which poses challenges in terms of generating, controlling, and managing a multitude of AAV vectors in every laboratory or company. This limitation in candidate exploration reduces the chances of discovering improved AAV variants. To address this issue, AAVnerGene has introduced the ATHENA I platform, enabling rapid and semi-high-throughput comparisons of AAV variants both in vivo and in vitro.
Design of ATHENA I
In the ATHENA I library, each capsid variant is associated with three different DNA-barcoded genomes, and all three of these genomes carry the same reporter gene. The use of three DNA barcodes for one capsid variant can minimize experimental variations and improve the accuracy of the results in high-throughput screening or selection processes.
In the standard ATHENA I kit, AAV genomes carried a CAG promoter drive EGFP expression, following by the Woodchuck hepatitis virus post-transcriptional regulatory element(WPRE) enhancer and a bovine growth hormone polyadenylation signal(bGH PolyA). The unique DNA barcode is located between the coding sequence and the PolyA signal. The CAG promoter is known for strong and ubiquitous expression in various cell types. The presence of the EGFP reporter gene allows for the enrichment of transgene-expressing cells, making it possible to identify and isolate cells that have taken up and expressed the AAV genomes. WPRE enhancer and bGH polyA can enhance EGFP expression and stabilize mRNA.
NGS technology is used to analyze the DNA barcode data. Researchers can use the unique DNA barcodes or transcribed RNA barcodes to assess the potential of different capsids for each target cell. It allows researchers to simultaneously assess hundreds of AAV vectors at once, both in vivo (in living organisms) and in vitro (in cell culture).
Products, Prices and Turnaround Times
AAVnerGene provides different premade ATHENA I AAV Capsid Libraries:
- AAV Capsid library(I)-Common: It contains 16 common used AAV serotypes, AAV1, AAV2, AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAVrh.10, AAV11, AAV12, AAV13, AAVrh74, AAV-DJ.
- AAV Capsid library(I)-tissues: Besides the 16 common AAV serotypes, it also contain 9 reported tissue-targeting AAV variants, with a total of 25 capsids.
Image Name Tags Price Buy hf:tags 
AAV Capsid Library(I)-BBB AAV-B1, AAV-F, AAV-MaCPNS1, AAV-PHP.C2, AAV-PHP.eB, AAV-PHP.S, AAV.Cap-B22, AAV.Cap-Mac, AAV/Olig001. $9,999.00 – $54,999.00 aav-b1 aav-f aav-macpns1 aav-php-c2 aav-php-eb aav-php-s aav-cap-b22 aav-cap-mac aav-olig001 AAV Capsid Library(I)-Common No additional tissue-targeting capsids $5,999.00 – $32,999.00 no-additional-tissue-targeting-capsids AAV Capsid Library(I)-Lung AAV2-ESGHGYF, AAV2-PRSTSDP, AAV2-Y444F-Y500F, AAV2.5T, AAV2H22, AAV5-PK2, AAV6.2, AAV6.2FF, AAV9.452sub.LUNG1 $9,999.00 – $54,999.00 aav2-esghgyf aav2-prstsdp aav2-y444f-y500f aav2-5t aav2h22 aav5-pk2 aav6-2 aav6-2ff aav9-452sub-lung1 AAV Capsid Library(I)-Retina AAV2-7M8, AAV2-Y444F-Y500F, AAV2.GL, AAV2.NN, AAV8-Y731F, AAV8BP2, AAVK9#12, AAVK9#4, ShH10 $9,999.00 – $49,999.00 aav2-7m8 aav2-y444f-y500f aav2-gl aav2-nn aav8-y731f aav8bp2 aavk912 aavk94 shh10
Highlights:
- The premade AAV Capsid Libraries(I) use CAG as promoter to drive EGFP and DNA barcodes.
- Presence of EGFP gene allows enrichment of transgene-expressing cells.
- Each capsid is associated with three different DNA-barcodes, to minimize experimental variations.
- Researchers can use the unique DNA barcodes or transcribed RNA barcodes, or both, to assess the potential of different capsids for each target cell.
- Each AAV capsid has a titer of approximately 1e12 GC in the premade AAV capsid libraries(I).
Notes.
1. AAVnerGene provides custom AAV capsid library(I) construction and production services. Customers can design their own expression cassettes and DNA barcode strategies, and add any AAV capsid into the libraries.
2. AAVnerGene has a capsid collection over 1000 different serotypes and variants.
Customers can choose their own AAV capsids, promoters, reporters, and other elements to make their own AAV capsid libraries(I). Please contact us for more information.
Design your reporter system: Choose a reporter gene or reporter cassette that can be easily detected or quantified to assess the transduction efficiency or tropism of AAV capsids. Commonly used reporter genes include fluorescent proteins like GFP or mCherry, or enzymatic reporters like luciferase or β-galactosidase.
Design your DNA barcode strategies: DNA barcodes are used to track and identify individual capsid variants within the library. Design a barcode strategy that allows for unique identification of each variant. This can be achieved by incorporating unique DNA barcode sequences into the capsid genomes, typically in non-coding regions. Consider the length and composition of the barcodes to ensure their uniqueness and compatibility with downstream sequencing or detection methods.
Select your capsids: Choose a set of capsids that represent a diverse range of serotypes or have different tropisms and transduction efficiencies. This can include well-characterized AAV serotypes or variants with known properties, as well as newly engineered capsids. Consider factors such as tissue specificity, receptor binding, and immunogenicity when selecting capsids for your library.
- Determine production scale: it’s important to consider the production scale needed to generate sufficient quantities of AAV vectors for downstream applications. The production scale will depend on factors such as the number of capsid variants in the library, the desired yield of each variant, and the intended use of the library.
Custom AAV capsid selection kits
AAVnerGene’s ATHENA I platform offers a comprehensive service for capsid library design and production, as well as one round of screening using next-generation sequencing (NGS). The screening can be performed in vitro using cell lines, primary cells and organoids, as well as in vivo using mouse and non-human primate (NHP) models. For specific screening needs, it is recommended to discuss with AAVnerGene’s technical support team to ensure the best results.
Potential models:
- In vitro: cell lines and primary cells, organoids
- In vivo: mouse and non-human primate (NHP)
