ATHENA AAV Capsid Screening Platform

ATHENA: AAVnerGene's Tissue-specific, Highly-transductive and Expressive New AAVs.

 

ATHENA is a two-step AAV selection Platform. First step, use our ATHENA I platform to select the best well known AAV vectors for each cell type. Second Step, use our high complexity, random peptide ATHENA II  AAV library to further optimize the selected capsids in ATHENA I.

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ATHENA I AAV Serotype Screening Platform

ATHENA I is a library that contains about 200 known capsids. Each capsid variant packages a DNA barcoded genome carrying a reporter gene(such us EGFP). The unique  barcode is located between coding sequence and Poly-A signal. Each capsid and barcoded transgene was paired and transfected separately. All the AAV vectors were harvested, purified and titerred, separately. Then, equal amount of each AAV vectors were pooled and ready to use. 

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ATHENA I is a small library that contains about 200 known capsids.  Each capsid variant packages a DNA barcoded genome carrying a reporter gene(such us EGFP). The unique  barcode is located between coding sequence and Poly-A signal. Each capsid and barcoded transgene was paired and transfected separately. All the AAV vectors were harvested, purified and titerred, separately. Then, equal amount of each AAV vectors were pooled and ready to use. 

Advantages of ATHENA I

  1. Presence of reporter gene allows enrichment of transgene-expressing cells and avoids the viral replication and packaging steps.

  2. The DNA barcodes screenings allow to evaluate the disruption of each capsids for each type of target cell.

  3. The transcribed RNA barcodes screenings allow the evaluating of putative capsids for each target cell. The efficacy of each AAV candidate is obtained by directly comparing the quantity of barcode before and after infection rather than comparing with other AAV variants.

  4. Hundreds of AAV vectors can be generated at once, pooled and tested together. The pre-made barcoded AAV pools allow us to fast and semi- high-throughput compare and select AAV variants both in vivo and in vitro.

Potential AAV capsids include, but not limit to:

 

Native AAV Serotypes: AAV1, AAV2, AAV3, AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAVrh10, AAV11, AAV12, AAV13.

  • Y/F mutants: single-, double-, triple-, quad-, penta-, hexa-,  mutation of any native serotypes.

  • Engineered AAV Capsids: AAV-PHP.B, AAV-PHP.eB, AAV-PHP.B2, AAV-PHP.B3, AAV-PHP.S, AAV-retro, AAV-7M8, AAV-F, AAV-Vec, AAV-Nec, AAV-SCH9, AAV-SCH2, AAV-SHH10, AAV6.1, AAV6.2, AAV6.2FF, AAV-B1, AAV-BP2, AAVv66, AAV-Anc110, AAV-Anc113, AAV-Anc126, AAV-Anc127, AAV-Anc80L65, AAV-Anc81, AAV-anc82, AAV-anc83, AAV-Anc84, etc.

  • Peptide Insert AAV2 vectors:

  • AAVs isolated  from Human: AAV-Hu.1, AAV-Hu.2, AAV-Hu.3, AAV-Hu.6, AAV-Hu.10, AAV-Hu.11, AAV-Hu.13, AAV-Hu.15, AAV-Hu.16, AAV-Hu.17, AAV-Hu.18, AAV-Hu.19, AAV-Hu.20,  AAV-Hu.37, AAV-Hu.45, AAV-Hu.47, AAV-Hu.48, AAV-Hu.49, AAV-Hu.52, AAV-Hu.58, etc.

  • AAV  isolated  from Rh: AAV-Rh10, AAV-Rh.22, AAV-Rh.23, AAV-Rh.24, AAV-Rh.32, AAV-Rh.32.33, AAV-Rh.33, AAV-Rh.34, AAV-Rh.35, AAV-Rh.39, AAV-Rh.43, AAV-Rh.47, AAV-Rh.49, AAV-Rh.52, AAV-Rh.74, etc.

  • AAVs isolated  from Rat: AAV-Rat-YY.12, AAV-Rat-YY.25, AAV-Rat-YY.78, AAV-Rat-YY.80, AAV-Rat-YY.90, AAV-Rat-YY.93, AAV-Rat-TT.54,AAV-Rat-HD.16, AAV-Rat-HD-20, AAV-Rat-HD-43, AAV-Rat-HD-94, AAV-Rat-MLP.26, AAV-Rat-MLP-6, AAV-Rat-XM.70,  etc.

  • AAVs isolated  from Other Specials: AAV-Po.1,AAV-Po.2,AAV-Po.3, AAV-Po.4, AAV-Po.5, AAV-Po.6, AAV-Po.7, AAV-Po.8, AAV-Go.1, AAV-VR355,AAV-Cy.2,AAV-Cy.3, AAV-Cy.4,AAV-Cy.5 AAV-Cy.6, AAV-Avian-DA, AAV-Avian-VR865, AAV-Bat-YNM, AAV-bovine,​ AAV-Pi.1,AAV-Pi.2,AAV-Pi.3, AAV-BB.1, AAV-BB.2,AAV-ch.5.

  • Peptide Insert AAV2 vectors: AAV2-588NGR, AAV2-MO7A, AAV2-MO7T, AAV2-MecA, AAV2-MecB, AAV2-rRGD587, AAV2-C4, AAV2-D10, AAV2-SIG, AAV2-MTP, AAV2-QPE, AAV2-VNT, AAV2-CNH, AAV2-CAP, AAV2-EYH, AAV2-587MTP,  etc

Custom AAV capsid selection kits

Our ATHENA I contains a EGFP expression cassette that driven by CAG promoter. Customers can choose their our promoters, reporters, AAV capsid and other elements to make their own AAV capsid selection kits. Please contact us for more information.

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Price and Time

Construction of backbone plasmids: 1-2 week.

Construction of random peptide library: 1-2 week

Production of AAV library: 1-2 weeks.  

Total time: 4~8 week.