| Field | Specification |
|---|---|
| Cellular Localization | |
| Form | Liquid |
| Product Type | |
| Promoter | |
| Reporter | |
| Storage |
Research background
This AAV is built for robust expression of a research payload in targeted cells or tissues. AAV-mediated delivery is widely used for stable transgene expression in vivo and in vitro.
Mechanism and expected readouts
Expression outcomes depend on promoter choice, cell state, delivery route, and the biology of the payload. Downstream readouts often include fluorescence imaging, histology, and functional assays matched to the encoded gene.
Expression design and interpretation
Expression is driven by a promoter choice that shapes cell-type bias and expression strength; human synapsin promoter, commonly used for broad neuronal expression. The expression design includes regulatory elements that can constrain activity to specific genetic contexts or experimental conditions. A built-in reporter/tag supports validation and localization; tdTomato provides a bright red fluorescent readout and is useful for sparse labeling and anatomy. The encoded payload is intended to support the stated experimental function (e.g., modulation, sensing, labeling, or control).
Subcellular targeting elements (when present) can bias localization and should be confirmed by imaging in your preparation.
Common research applications
- Cell labeling and anatomical tracing (when paired with appropriate reporters)
- Overexpression or rescue experiments for mechanistic studies
- Tool-building workflows combining expression with physiology or behavior
Experimental considerations
- Plan expression time (often weeks in vivo) according to your tissue and readout
- Validate targeting and expression pattern in pilot experiments
- Include appropriate controls (reporter-only, inactive variants, or sham injection)
Controls and validation
Typical validation includes confirming expression pattern and level, verifying functional activity with an assay matched to the payload (e.g., imaging, electrophysiology, pharmacology, or behavior), and using appropriate negative controls.
At present, the main purification approaches for rAAV include:
- Ultracentrifugation density-gradient methods, using cesium chloride (CsCl) or iodixanol as the gradient medium;
- Chemical reagent precipitation/extraction methods, mainly using PEG, ammonium sulfate, chloroform, etc.;
- Chromatographic purification methods, primarily based on affinity and ion-exchange principles.
Depending on customers’ different application needs, we can integrate multiple methods to produce high-titer, high-purity, high-quality rAAV viral products.
Titer Determination
Quantitative PCR (qPCR) is used to measure the copy number of the rAAV genome, reported in vg/mL (vector genomes per mL).
Figure 1 Sample Amplification Curve
Figure 2 Standard Curve
Purity Testing
rAAV purity is assessed by protein gel electrophoresis (SDS-PAGE) to evaluate the content of rAAV capsid proteins, typically expressed as a percentage (%).
The AAV capsid is composed of three structural proteins: VP1 (~87 kDa) VP2 (~73 kDa) VP3 (~62 kDa) Therefore, three distinct bands will appear on the SDS-PAGE gel image.
Figure 3 SDS-PAGE analysis shows rAAV purity is ≥97%
Quality Assurance: We ensure that the data and materials provided to customers are truthful and reliable.
To choose a serotype, start with your target tissue and delivery route, then pick a capsid with a proven track record in that setting and benchmark 1–3 alternatives. For liver-directed systemic expression, AAV8 is a common first choice; for broad systemic delivery and strong cardiac performance, AAV9 is often preferred. For local CNS injections, AAV2 (or AAV1/2 for improved spread) is a reliable starting point, while rAAV2-retro is ideal when you need retrograde labeling from projection targets. For mouse brain-wide delivery by IV, AAV-PHP.eB is frequently used, and AAV-PHP.S is often selected for PNS/DRG-enriched programs. For muscle, AAV1, AAV6 (and AAV6 variants) or MyoAAV 2A are common starting points; for airway/lung, AAV5 or AAV6 are typical benchmarks. For ocular work, route matters: AAV-7m8 is often chosen for intravitreal retinal delivery, while AAV2/AAV5/Anc80L65 are common comparators depending on the target layer. If you’re unsure or working in a new model, include a broad performer like AAV-DJ and/or a small screening panel (e.g., PAN/BI30/SCH9) to quickly identify the best capsid in your exact system.
| Serotype / Capsid | Best-known strengths | Common applications (examples) |
|---|---|---|
| AAV2/1 | High efficiency in skeletal muscle | Muscle gene expression, local delivery to muscle; neuromuscular research |
| AAV2/2 | Reliable neuronal transduction with local CNS injection; strong history of use | Local brain/spinal injections; retinal subretinal delivery; general benchmarking |
| AAV1/2 | Hybrid capsid often used for enhanced neuronal transduction/spread | CNS gene expression where improved diffusion vs AAV2 is desired |
| AAV2/5 | Strong performance in select CNS and airway contexts | CNS research; airway/respiratory delivery; retinal programs (route-dependent) |
| AAV2/6 | Efficient in muscle and airway/lung | Muscle delivery; pulmonary/airway delivery; selected ex vivo workflows |
| AAV2/6m | AAV6 variant for enhanced entry in muscle/airway settings | Higher-efficiency muscle or airway delivery (model-dependent) |
| AAV2/6.2 | AAV6 variant used to boost muscle/airway delivery | Muscle and respiratory delivery where AAV6 is a baseline |
| AAV2/8 | High efficiency in liver | Liver-directed gene expression; systemic dosing where liver is the primary target |
| AAV2/9 | Broad systemic delivery; strong in heart and useful for CNS access in rodents | Cardiac studies; systemic delivery; CNS programs (route/age dependent) |
| rAAV2-retro | Retrograde transport in neuronal circuits | Circuit mapping; projection-based targeting; retrograde labeling from injection sites |
| AAV2-PHP.eB | Very high whole-CNS transduction in mice (systemic) | Mouse brain-wide delivery for neuroscience and neurogenetics |
| AAV2-B10 | Engineered capsid used in targeting/optimization workflows | Tissue-targeting exploration and comparative capsid testing |
| AAV2-PHP.S | Enhanced peripheral nervous system (PNS) targeting in rodents | DRG/PNS studies; peripheral neurobiology (systemic delivery) |
| AAV2-PAN | Designed for broad (“pan”) transduction in some settings | Broad expression screens; programs needing wide tissue coverage |
| AAV2-DJ | Robust, broad transduction; strong general-purpose performer | High-efficiency transduction in many cell types; discovery/validation work |
| AAV2-7m8 | Optimized for retinal delivery via intravitreal injection | Retina gene delivery with intravitreal route; ocular research |
| AAV2-ShH10 | Retinal tropism with reported preference in retinal glia | Retinal biology; Müller glia–focused programs (route dependent) |
| AAV2-Rh10 | Frequently used for CNS applications | CNS delivery (local routes); neuroscience studies |
| AAV2-Anc80L65 | Broad performance; widely used in sensory/inner ear and ocular settings | Inner ear studies; ocular programs; broad benchmarking |
| AAV2-BR1 | Enrichment for brain vasculature/endothelium | BBB/endothelial targeting; neurovascular research |
| AAV2-BI30 | Engineered capsid for targeting/optimization panels | Capsid screening; tissue targeting exploration |
| AAV2-SCH9 | Engineered capsid for targeting/optimization panels | Capsid screening; comparative tissue targeting |
| MaCPNS1 | Candidate capsid in neural delivery panels | CNS/PNS capsid comparisons; screening and optimization |
| MaCPNS2 | Candidate capsid in neural delivery panels | CNS/PNS capsid comparisons; screening and optimization |
| mac | Candidate capsid in broad screening panels | Capsid screening and optimization across routes/models |
| AAV2/11 | Used for airway/epithelial delivery in some programs | Respiratory/airway targeting; comparative testing vs AAV5/6 |
| MyoAAV 2A | Optimized for skeletal muscle targeting | Systemic or local muscle delivery; myopathy/neuromuscular studies |
| VCAP-102 | Candidate capsid used for vascular/endothelial-focused programs | Vascular biology; endothelial targeting studies |
Can’t find the AAV you need—or require a custom design and packaging service? We offer end-to-end support for diverse research and therapeutic needs, including vector design and cloning, AAV packaging services (serotype/capsid selection and production), QC & characterization (project-appropriate testing and documentation), and library preparation for pooled or library-style workflows (project dependent). Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
References
- Boyden ES, Zhang F, Bamberg E, Nagel G, Deisseroth K (2005). Millisecond-timescale, genetically targeted optical control of neural activity. Nat Neurosci. DOI: 10.1038/nn1525 [PMID: 16116447]. Original ChR2 optogenetics paper
- Fenno L, Yizhar O, Deisseroth K (2011). Optogenetics in neural systems. Neuron. DOI: 10.1016/j.neuron.2011.06.004 [PMID: 21867882]. Comprehensive review of ChR2 variants and optogenetic tools
- Shaner NC, Campbell RE, Steinbach PA, et al. (2004). Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol. DOI: 10.1038/nbt1037 [PMID: 15558047]. tdTomato tandem dimer fluorescent protein
- Kügler S, Kilic E, Bähr M (2003). Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression from an adenoviral vector in the adult rat brain. Gene Ther. DOI: 10.1038/sj.gt.3301905 [PMID: 12595892]. hSyn promoter — pan-neuronal specificity
- Ayuso E, Mingozzi F, Montane J, et al. (2010). Production and purification of serotype 1, 2, and 5 recombinant adeno-associated viral vectors. Gene Ther. DOI: 10.1038/gt.2010.17 [PMID: 20237510]. Sf9/baculovirus AAV production system
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