| Field | Specification |
|---|---|
| Cellular Localization | |
| Form | Liquid |
| Function | |
| Product Type | |
| Promoter | |
| Reporter | |
| Storage |
Research background
This AAV provides recombinase-based genetic access, enabling conditional control of downstream payloads in genetically defined cells. Recombinase tools are widely used to restrict expression, create intersectional logic, or enable lineage/cell-type targeting.
Mechanism and expected readouts
Recombinase expression catalyzes site-specific DNA recombination at matching target sites, permanently switching genetic states in infected cells. This can gate expression of FLEX/DIO payloads or enable intersectional strategies when combined with other recombinases.
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 construct includes regulatory logic that controls where/when the payload is active; Cre-dependent control restricts expression to cells expressing Cre recombinase. A built-in reporter/tag supports validation and localization; GFP/EGFP provides a green fluorescent readout for expression, morphology, and targeting validation. 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
- Conditional targeting for cell-type or projection-specific experiments
- Intersectional labeling or manipulation strategies
- Permanent genetic switching for lineage or fate-mapping workflows
Experimental considerations
- Verify recombination efficiency with a compatible reporter in your system
- Account for irreversible switching when designing longitudinal studies
- Use appropriate negative controls (no recombinase or mismatched recombinase lines)
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
- Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994). Green fluorescent protein as a marker for gene expression. Science. DOI: 10.1126/science.8303295 [PMID: 8303295]. Original GFP/EGFP reporter paper
- 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
- Gradinaru V, Thompson KR, Zhang F, et al. (2007). Targeting and readout strategies for fast optical neural control in vitro and in vivo. J Neurosci. DOI: 10.1523/JNEUROSCI.3606-07.2007 [PMID: 17959775]. Double-floxed inverted open reading frame (DIO) design for Cre-dependent AAV
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