AAV Library Services
AAVnerGene offers comprehensive AAV library solutions—from custom AAV library design to high efficient AAV library construction and high-quality AAV library production, to directed evolution services. Our platform supports a diverse range of AAV library types tailored for discovery, optimization, and functional screening. Our core offerings include:
With end-to-end capabilities and deep expertise in AAV engineering, AAVnerGene empowers researchers to accelerate innovation and uncover next-generation AAV vectors by providing the following AAV library services:
- AAV Library Design/Construction
- AAV Library Packaging/Production
- AAV Library Screening/Evolution
AAV capsids possess nine surface-exposed variable regions (VRs), VR-I to VR-IX, essential for determining tropism and receptor binding. Current directed evolution efforts primarily focus on VR-VIII and VR-IV. VR-VIII is particularly the spike positions around the 3-fold axis (N587/R588 of AAV2 or Q588/A589 of AAV9), which serve as key engineering sites. This focus has led to well-known evolved capsids like AAV2-7M8, AAV-PhP.eB, AAV-Cap.Mac, and MyoAAVs. VR-IV, the most radially protruding loop, plays a role in neutralizing antibody and surface receptor binding due to its proximity to the 3-fold symmetry axis. Compared to VR-VIII, VR-IV is less permissive for peptide inserts. However, enhancing diversity in VR-IV can improve interactions with existing VR-VIII mutations and refine transduction, exemplified by AAV.Cap.B22, engineered from AAV-PHP.eB. Notably, the evolution of the AAV6 VR-IV library alone produced Ark313, which shows high transduction efficiency in murine T cells.
To date, few engineering efforts have targeted other VRs, but they contribute to local topological differences among AAVs. Specifically:
- VR-V, along with VR-IV and VR-VIII, forms the top of the 3-fold protrusion.
- VR-II constitutes the top of the 5-fold channel.
- VR-VI and VR-VII form the base of the 3-fold protrusion.
- VR-I, VR-III, VR-VII, and VR-IX contribute to the 2/5-fold wall.
These VRs also dictate functional differences, influencing receptor attachment, transduction efficiency, and antigenic reactivity among AAVs.
To evolve capsids through multiple rounds of selection, an AAV expression cassette is essential for both production and selection. This cassette typically includes the necessary genetic elements to package the AAV capsid library, produce functional viral particles, and enable the identification of successful variants. Key components of the expression cassette often include:
- ITRs (Inverted Terminal Repeats): Flanking the expression cassette, ITRs are essential for AAV genome packaging and replication.
- Promoters: The choice of promoter is critical, as capsid proteins need to be produced in the correct ratio during the packaging step and expressed at high levels during the selection steps. A hybrid promoter is commonly used to achieve this purpose. A strong or tissue-specific promoter is often employed to drive capsid gene expression during selection, while the P40 promoter is used to ensure proper capsid assembly.
- Capsid Gene with Variable Regions: The AAV capsid gene is modified to incorporate random peptide insertions or mutations in its variable regions (VRs), enabling the generation of a diverse library of capsid variants. These modifications allow for the exploration of new tropisms, transduction efficiencies, and immunogenic profiles.
At AAVnerGene, we empower researchers to accelerate AAV capsid discovery through our advanced AAV Barcode Capsid Library Production Services. Barcode-Seq utilizes deep-sequencing technologies such as Illumina amplicon-seq to track the performance of individual AAV vector within complex mixtures. In this approach, each AAV capsid variant encapsulates an identical transgene expression construct that differs only by a short, unique DNA barcode. This barcode serves as a molecular identifier, allowing precise quantification of each variant’s abundance and transduction efficiency across different tissues or cell types. To minimize potential barcode-dependent bias, each variant can be linked to multiple independent barcodes, ensuring statistically robust and reproducible results. Designed for capsid evolution and tissue tropism studies, these kits enable parallel evaluation of hundreds or thousands of AAV capsid variants in vivo or in vitro. Researchers can determine which variants show superior packaging efficiency, transduction, or tissue specificity — all in a single sequencing run.
AAVnerGene provides specialized AAV promoter library services aimed at accelerating the discovery and optimization of regulatory elements for precise and efficient gene expression. Leveraging advanced DNA barcode technology, our comprehensive platform allows researchers to screen and identify promoters that drive tissue-specific, cell-type–specific, or inducible transgene expression through AAV vectors.
Our Barcode-Seq–based platform offers:
High-Throughput Screening: Rapid, simultaneous evaluation of multiple promoters and enhancers to pinpoint the most effective regulatory elements.
Precision and Accuracy: Quantitative measurement of transgene expression levels and specificity within target cells or tissues.
Data-Driven Optimization: In-depth analysis to select the ideal cis-regulatory elements tailored to your gene therapy goals.
Similar to our AAV Promoter Library, the AAV Enhancer Library also utilizes our Barcode-Seq technology to enable high-throughput and quantitative screening of enhancer elements. Each enhancer is assigned to multiple unique DNA barcodes, ensuring robust statistical power, reduced bias, and highly reliable performance measurements across complex biological systems.
AAVnerGene provides end-to-end AAV shRNA library services designed for large-scale functional screening in vitro and in vivo. Leveraging our proprietary AAVone®, AAVdual and AAVtri production systems, and high-diversity capsid library expertise, we deliver uniform, high-quality RNA-interference libraries tailored to your research needs.
AAVnerGene provides comprehensive, high-quality AAV sgRNA library services designed to accelerate functional genomics, gene knockout screening, and CRISPR-based discovery workflows. Our platform integrates optimized AAV vector engineering, precise library synthesis, and high-yield viral packaging to deliver ready-to-use sgRNA libraries tailored to your research needs.
We generate sgRNA libraries targeting whole genomes, specific pathways, gene families, or user-defined gene sets. Libraries can be designed for CRISPR-Cas9, Cas12, or other nuclease systems.
Using advanced oligo synthesis and cloning strategies, we ensure uniform representation, low bias, and high library diversity. Each library undergoes deep sequencing QC to confirm coverage and integrity.
AAVnerGene specializes in packaging large, complex sgRNA libraries using our proprietary mini-pHelper-Rep plasmid and high-efficiency production systems (AAVone/AAVone2.0). This enables:
Superior diversity retention
High genome copies per cell
Enhanced full-particle ratios
Reliable, scalable viral yields
Libraries can be packaged into a wide range of AAV capsids, including AAV9, AAV6, AAV-PHP variants, engineered capsids, or customer-provided serotypes—enabling tissue-specific or systemic delivery in vitro or in vivo.