Loading ....
Integrated AAV Offerings: From Cutting-Edge Products to Advanced Custom Services
Overcoming AAV Technology Challenges with Expertly Tailored Vector Design and Production
ATHENA I AAV Capsid Platform
The ATHENA I AAV Capsid Platform is an advanced triple-barcode capsid library developed by our partner, AAVnerGene, designed to systematically evaluate and optimize AAV serotypes and variants for a wide range of gene delivery and therapeutic applications. Leveraging Barcode-Seq technology, this platform provides a powerful, semi-high-throughput method for assessing the efficiency and suitability of over 1,000 distinct AAV capsids both in vivo (within living organisms) and in vitro (in cell culture).
Challenges in AAV Vector Development
AAV vectors have become crucial tools for gene delivery and gene therapy, thanks to their ability to target specific cell types and tissues. However, with the discovery of numerous natural AAV serotypes and the increasing number of engineered variants, the challenge remains to identify the most effective capsids tailored for specific therapeutic applications. Traditional methods involve testing AAV variants individually, which is cumbersome and inefficient, especially when handling a large number of variants. This limitation hinders the ability to explore a broad range of candidates and discover optimized AAV capsids.
Purpose and Advantages of the ATHENA I Platform
The ATHENA I platform addresses these challenges by enabling researchers to perform rapid and semi-high-throughput comparisons of AAV variants. Each capsid variant in the ATHENA I library is associated with three unique DNA-barcoded genomes, all carrying the same reporter gene. This triple-barcode system minimizes experimental variations and enhances the accuracy of high-throughput screening, allowing researchers to determine the most efficient AAV capsid variant for their specific needs.
Design and Components of the ATHENA I Platform
In the standard ATHENA I kit, AAV genomes are driven by a CAG promoter that ensures strong and ubiquitous expression across various cell types. The EGFP reporter gene allows for the identification and enrichment of transgene-expressing cells, facilitating the selection of cells that have successfully taken up and expressed the AAV genomes. The inclusion of the Woodchuck Hepatitis Virus Post-Transcriptional Regulatory Element (WPRE) enhancer and the bovine growth hormone polyadenylation signal (bGH PolyA) further enhances EGFP expression and stabilizes mRNA, ensuring robust and reliable results.
High-Throughput Screening with NGS Technology
Utilizing Next-Generation Sequencing (NGS) technology, the ATHENA I platform allows researchers to analyze DNA or RNA barcode data to evaluate the potential of different capsids for specific target cells. This high-throughput approach enables the simultaneous assessment of hundreds of AAV vectors, significantly streamlining the process of capsid optimization and accelerating the discovery of next-generation AAV vectors with improved properties.
AAV Capsids Barcode Kits
AAV Capsid Barcode Kit - Common: This kit includes 16 widely used AAV serotypes: AAV1, AAV2, AAV2-Retro, AAV3B, AAV5, AAV6, AAV7, AAV8, AAV9, AAV11, AAV12, AAV13, AAVrh.10, AAVrh.74, AAV-DJ, and AAV4-Y729F. It is ideal for broad-spectrum AAV testing and screening applications.
| SKU | Name | Serotypes | Scales |
|---|---|---|---|
| DA005001 | AAV Capsid Barcode Kit-Common CAG-EGFP |
16 common | 1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
| DA001006 | AAV Capsid Barcode Kit-Common CMV-mCherry |
16 common | 1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
| DA023006 | AAV Capsid Barcode Kit-Common sc-CMV-mCherry |
16 common | 1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
AAV Capsid Barcode Kit - Tissue-specific: In addition to the 16 common AAV serotypes, this kit contains 9 additional tissue-targeting AAV variants, totaling 24 capsids. It is specifically designed for researchers focusing on tissue-specific applications and targeting. The AAV cassette used for tissue specific kits is CAG-EGFP.
| SKU | Name | Serotypes | Scale |
|---|---|---|---|
| DH005001B | AAV Capsid Barcode Kit-BBB | 16 common + 9 BBB targeted capsids: AAV-B1, AAV-F, AAV-MaCPNS1, AAV-PHP.C2, AAV-PHP.eB, AAV-PHP.S, AAV.Cap-B22, AAV.Cap-Mac, AAV/Olig001. |
1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
| DH005001L | AAV Capsid Barcode Kit-Lung | 16 common + 9 lung targeted capsids: AAV2-ESGHGYF, AAV2-PRSTSDP, AAV2-Y444F-Y500F, AAV2.5T, AAV2H22, AAV5-PK2, AAV6.2, AAV6.2FF, AAV9.452sub.LUNG1 |
1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
| DH005001M | AAV Capsid Barcode Kit-Myo | 16 common + 9 Myo targeted capsids: AAV587MTP, AAV9.45, AAV9.61, AAVM41, AAVMYO, AAVmyo2, AAVmyo3, myoAAV-2A, myoAAV-4A |
1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
| DH005001R | AAV Capsid Barcode Kit-Retina | 16 common + 9 retina targeted capsids: AAV2-7M8, AAV2-Y444F-Y500F, AAV2.GL, AAV2.NN, AAV8-Y731F, AAV8BP2, AAVK9#12, AAVK9#4, ShH10 |
1x10^12 VG/Capsid 2x10^12 VG/Capsid 5x10^12 VG/Capsid 10x10^12 VG/Capsid |
AAV Serotype Testing Kits
AAV Serotype Testing Kits provide a quicker method to identify a specific subset of AAV serotypes by measuring the expression levels of fluorescent proteins (EGFP, mCherry, TdTomato) or luciferases (Gluc, Rluc, Fluc, Cluc). This approach is particularly useful when researchers have a clear understanding of the target cell lines or tissues and seek a streamlined way to narrow down their choices to a few optimal AAV serotypes.
In this category, 15 most used AAV serotypes (AAV1, AAV2, AAV3B, AAV5, AAV6, AAV7, AAV8, AAV9, AAV11, AAV12, AAV13, AAV-DJ, AAVrh.10, AAVrh.74, and AAV2-Retro) are included in each kit. The vectors carry fluorescent protein (EGFP, mCherry, TdTomato) or reporter luciferase (Fluc, Rluc, Cluc, Gluc), which is driven by a promoter (CAG, CMV, EF1a or hSyn). The kits are pretended to provide useful tools for customers, with minimum cost, in testing AAV transduction efficiency, testing tissue specific tropism related to serotypes, and using as negative control to examine whether the observed biological effects come from specific transgenes. We also provide custom AAV Capsid Kits developing services.
| SKU | Product Name | Genome | Scales |
|---|---|---|---|
| PK0001 | AAV Serotype Testing Kit-CAG-EGFP | AAV-CAG-EGFP | 50ul x 1E+13 GC/mL |
| PK0002 | AAV Serotype Testing Kit-CAG-mCherry | AAV-CAG-mCherry | 50ul x 1E+13 GC/mL |
| PK0003 | AAV Serotype Testing Kit-CMV-mCherry |
AAV-CMV-mCherry | 50ul x 1E+13 GC/mL |
| PK0004 | AAV Serotype Testing Kit-CAG-TdTomato | AAV-CAG-TdTomato | 50ul x 1E+13 GC/mL |
| PK0005 | AAV Serotype Testing Kit-CMV-Fluc | AAV-CMV-Fluc | 50ul x 1E+13 GC/mL |
| PK0006 | AAV Serotype Testing Kit-EF1a-EGFP | AAV-EF1a-EGFP | 50ul x 1E+13 GC/mL |
| PK0007 | AAV Serotype Testing Kit-EF1a-mCherry | AAV-EF1a-mCherry | 50ul x 1E+13 GC/mL |
| PK0009 | AAV Serotype Testing Kit-hSyn-mCherry | AAV-hSyn-mCherry | 50ul x 1E+13 GC/mL |
Custom AAV Capsid Barcode Library Design
1. Select Your Promoter: Choose from a range of promoters such as CMV, CAG, EF1a, Syn, TTR, CaMKII, hAAT, or other options to drive the expression of your gene of interest.
2. Select Your Reporter: Select a reporter gene that can be easily detected or quantified to assess the transduction efficiency or tropism of AAV capsids. Commonly used reporter genes include EGFP, mCherry, TdTomato, Gluc, Fluc, Rluc, Cluc, AAT, LacZ, among others.
3. Select Your Capsid: Choose from a diverse range of AAV serotypes or engineered variants such as AAV1, AAV2, AAV3B, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, and others. Consider factors such as tissue specificity, receptor binding, and immunogenicity for your library.
4. Contact Us: Reach out to our team at [email protected] to discuss your specific requirements and initiate the custom library creation process.
5. Make Barcoded Reporter Library: Insert random barcodes between reporters and PolyA to create a barcoded library. Miniprep of plasmids and sequencing are used to confirm the barcode in each plasmid.
6. Produce AAV Capsid Library: Each capsid is matched to one unique barcoded plasmid. AAV vectors are produced separately, then pooled and purified together. qPCR primers are designed according to the barcodes.
7. Perform Experiments: Conduct screening experiments in vitro using cell lines, primary cells, or organoids, or in vivo using mouse and non-human primate (NHP) models. Collect Data: Use qPCR, NGS, and data analysis to evaluate the performance and efficiency of the AAV capsid variants.
Custom AAV Capsid Selection Kits
AAVnerGene’s ATHENA I platform
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). This screening can be performed both in vitro (e.g., cell lines, primary cells, organoids) and in vivo (e.g., mouse and NHP models). For specific screening needs, we recommend discussing them with AAVnerGene’s technical support team to ensure the best results.
Potential models:
In vitro: cell lines, primary cells, organoids
In vivo: mouse and non-human primate (NHP)
Contact Us
AAV Services
AAV Vector Design & Cloning
Expert consultation, design, gene synthesis, and cloning of custom AAV vectors tailored to your research needs.
Learn moreAAV Packaging Service
Reliable, affordable, and fully customizable AAV packaging service with diverse serotype options and high titer.
Learn moreAAV QC & Characterization
Comprehensive quality control assays including qPCR, SDS-PAGE, mass photometry, and next-generation sequencing.
Learn moreOther Viruses at Biohippo
Neural Circuit Tracing
Biohippo offers a variety of viral vectors, including RV, HSV, PRV, and VSV, to replace conventional tracers with the highly efficient, more specific, and less invasive viral method.
Learn More
Lentivirus
Biohippo offers TF reporter lentivirus, immunotherapy lentivirus, validated shRNA, cDNA, and miRNA lentiviruses to meet diverse research needs. We also offer customized service at affodable price.
Learn More