| Field | Specification |
|---|---|
| Form | Liquid |
| Function | |
| Plasmid Backbone | |
| Product Type | |
| Production System | |
| Promoter | |
| Reporter | |
| Storage |
Overview
AAV-GFP (AAV serotype 1) AAV (AAV1-GFP) is an AAV vector packaged in AAV1 under the CMV promoter that delivers eGFP to mammalian cells. Researchers commonly use this vector for reporter assay; in vivo gene delivery; cell labeling.
Key elements and design rationale
- Capsid (serotype): AAV1. broad transduction with notable activity in skeletal muscle, central nervous system (neurons), and retina.
- Promoter: CMV — human cytomegalovirus immediate-early promoter; strong, broadly active in most mammalian cell types.
- Payload: eGFP — enhanced green fluorescent protein; standard fluorescent reporter.
- Genome backbone: Recombinant AAV (single-stranded unless explicitly noted as scAAV) flanked by AAV2 ITRs.
Biological background
eGFP is an enhanced variant of green fluorescent protein originally cloned from Aequorea victoria. It folds autonomously in mammalian cells and produces a chromophore by cyclization of an internal Ser-Tyr-Gly tripeptide. eGFP excitation and emission peaks (~488 nm excitation / ~507 nm emission) are well-matched to standard fluorescence microscopy and flow cytometry hardware.
Because eGFP signal is proportional (within limits) to expression level, eGFP-expressing AAVs are commonly used as reporters of transduction efficiency, promoter activity, and targeting fidelity.
The CMV promoter — human cytomegalovirus immediate-early promoter; strong, broadly active in most mammalian cell types — drives expression of the payload from the AAV cassette in this product. Promoter–capsid combinations together determine where and at what level the payload is expressed.
Research relevance and current trends
- Reporter AAVs continue to be standard tools for benchmarking new capsid variants and engineered serotypes (e.g., directed evolution capsids, MyoAAV-class capsids, AAV.PHP-family capsids in mice).
- Single-cell readouts (e.g., scRNA-seq, FACS) increasingly use reporter AAVs to define transduced populations within heterogeneous tissue.
- AAV vector engineering — including capsid evolution, capsid shuffling, and rational design — continues to expand the spectrum of accessible tissues and cell types.
Common research applications
- Tracking transduction efficiency across cell types or tissue regions.
- Benchmarking new capsids or promoters in vitro and in vivo.
- Acting as a labeling vector in dual-color experiments.
Use this product within experimental designs that include matched controls (capsid, promoter, dose, route) and a transduction validation step before interpreting payload-specific phenotypes.
Notes for experimental interpretation
- Confirm transduction efficiency in the target cell population before drawing payload-specific conclusions; reporter signal alone validates only that the vector reached and expressed in the cells.
- Match AAV dose, capsid, promoter, and route across all conditions when comparing payload to control; differences in any of these confound payload-specific interpretation.
- Avoid repeated freeze–thaw cycles of AAV stocks — aliquot upon first thaw.
- AAV biology, including tropism, can differ between species, strains, ages, and routes — confirm in your specific system.
Choose an AAV capsid based on your target tissue/cell type and delivery route, then benchmark 1–2 alternative serotypes empirically. The capsid (serotype) determines surface attachment and uptake; the cassette and promoter then control where and how strongly expression occurs once cells are transduced. The reference table below summarizes well-established tropism patterns — actual transduction efficiency depends on cell type, route, dose, anti-AAV neutralizing antibodies, and species.
Serotype × tissue tropism reference
| Serotype | Primary attachment / receptor | Best-supported tissues / cells | Common use cases |
|---|---|---|---|
| AAV1 | α-2,3 / α-2,6 N-linked sialic acid | Skeletal muscle, cardiac muscle, CNS neurons, retinal pigment epithelium | Intramuscular and stereotaxic CNS injection; broad neuronal labeling |
| AAV2 | Heparan sulfate proteoglycan (HSPG); coreceptors FGFR1, HGFR | CNS neurons, retinal ganglion cells, kidney, vascular smooth muscle | Stereotaxic CNS injection; intravitreal eye delivery; standard CNS workhorse |
| AAV4 | α-2,3 O-linked sialic acid | Retinal pigment epithelium, ependymal cells of brain ventricles | Subretinal RPE labeling; intracerebroventricular ependyma transduction |
| AAV5 | α-2,3 N-linked sialic acid; PDGFR coreceptor | Airway epithelium, CNS (astrocytes prominent), retinal photoreceptors | Intratracheal lung delivery; CNS astrocyte transduction; subretinal photoreceptor |
| AAV6 | Sialic acid + HSPG; EGFR coreceptor | Skeletal muscle, cardiac muscle, lung, hematopoietic cells (incl. T cells, HSPCs) | Intramuscular delivery; ex vivo HSPC engineering; intratracheal lung |
| AAV8 | 37/67 kDa Laminin receptor (LamR) | Liver (hepatocytes), cardiac muscle, skeletal muscle, retina, pancreas | Systemic IV → liver-directed expression (gold standard); cardiac and pancreatic |
| AAV9 | Terminal N-linked galactose; LamR | Cardiac muscle, skeletal muscle, CNS (crosses BBB in neonates and at high IV dose), liver, lung | Systemic IV for cardiac/skeletal muscle and CNS; intrathecal for spinal cord and DRG |
| AAV-DJ | Engineered chimera (directed evolution from AAV2/8/9) | Broad efficient transduction of mammalian cell lines and primary cells in vitro | In vitro transduction where high efficiency across cell lines is needed; not intended for systemic in vivo use (rapid clearance) |
Selection workflow
- Define the readout. Identify your target tissue/cell type and the experimental window (acute days, weeks, or chronic months).
- Match capsid to tissue. Use the table above as a starting point. For systemic IV, AAV8 (liver), AAV9 (cardiac/skeletal muscle, CNS via BBB), and AAV6 (muscle/lung) are the most common choices. For stereotaxic CNS, AAV2 / AAV5 / AAV9 are first-line. For skeletal muscle, AAV1 / AAV6 / AAV8 / AAV9 all perform well with subtle tissue and species differences.
- Match promoter to expression goal. CMV / CAG / CBA give strong, broadly active expression. Cell-type-specific promoters (CamKIIα, hSyn, GFAP, cTNT, αMHC, TBG, Ttr) restrict expression even when the capsid transduces multiple populations. Capsid-restricted tropism and promoter-restricted expression are independent layers of specificity that can be combined.
- Run a small dose-response. In vitro, test a 10× MOI range with a reporter AAV (e.g., AAV-GFP) of the same serotype to fix optimal MOI before switching to your transgene. In vivo, pilot 2–3 doses with a reporter or matched control vector before scaling.
- Use proper controls. Match capsid serotype, promoter, and dose between test and control vectors. Empty / Null capsid controls (e.g., AAV-Null) match for capsid- and dose-related effects independent of payload; LacZ or GFP-only vectors match for transgene-expression load.
Practical considerations
- Anti-capsid neutralizing antibodies. Pre-existing immunity against AAV2 and several other serotypes is common in human and primate studies and reduces transduction. This is less of a concern in inbred laboratory mouse strains but is reportable in NHP and human-relevant work.
- Route matters as much as capsid. The same capsid can give very different tropism by intravenous vs. intramuscular vs. intrathecal vs. stereotaxic vs. subretinal injection. The "best" capsid for a tissue is route-specific.
- Single-stranded vs. self-complementary (scAAV). Standard recombinant AAV is single-stranded and requires second-strand synthesis after entry, leading to a 1–3 week onset to peak expression. scAAV bypasses this step (faster onset, ~3–7 days) at the cost of half the packaging capacity (~2.4 kb vs. ~4.7 kb).
- ITR backbone. Nearly all recombinant AAVs — across capsid serotypes — use AAV2 ITRs. The capsid identity and the ITR identity are independent design choices.
- Empirical validation is required. Tropism summaries are starting points. Final serotype selection should be validated in a pilot experiment in your specific cell line, animal model, and route of administration.
Selected references on AAV biology and tropism: Wu Z, Asokan A, Samulski RJ. Adeno-associated virus serotypes: vector toolkit for human gene therapy. Mol Ther 2006;14(3):316–327. Zincarelli C, Soltys S, Rengo G, Rabinowitz JE. Analysis of AAV serotypes 1–9 mediated gene expression and tropism in mice after systemic injection. Mol Ther 2008;16(6):1073–1080. Srivastava A. In vivo tissue-tropism of adeno-associated viral vectors. Curr Opin Virol 2016;21:75–80. Pillay S, et al. An essential receptor for adeno-associated virus infection. Nature 2016;530:108–112.
What is this AAV product, briefly?
How should this AAV be stored and handled upon receipt?
What MOI should I start with?
What tropism should I expect from AAV1?
What controls should I include alongside this AAV?
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Selected References
- Gombash SE, Manfredsson FP, Mandel RJ, et al. Neuroprotective potential of pleiotrophin overexpression in the striatonigral pathway compared with overexpression in both the striatonigral and nigrostriatal pathways. Gene Ther 2014. PMID: 24807806
- Rovó Z, Mátyás F, Barthó P, et al. Phasic, nonsynaptic GABA-A receptor-mediated inhibition entrains thalamocortical oscillations. J Neurosci 2014. PMID: 24849349
- Du K, Zheng S, Zhang Q, et al. Pten Deletion Promotes Regrowth of Corticospinal Tract Axons 1 Year after Spinal Cord Injury. J Neurosci 2015. PMID: 26134657
- Das S, Morvan F, Morozzi G, et al. ATP Citrate Lyase Regulates Myofiber Differentiation and Increases Regeneration by Altering Histone Acetylation. Cell Rep 2017. PMID: 29241530
- Functional aspects of meningeal lymphatics in ageing and Alzheimer’s disease
- Hammond SL, Leek AN, Richman EH, et al. Cellular selectivity of AAV serotypes for gene delivery in neurons and astrocytes by neonatal intracerebroventricular injection. PLoS One 2017. PMID: 29244806
- Escalated Alcohol Self-Administration and Sensitivity to Yohimbine-Induced Reinstatement in Alcohol Preferring Rats: Potential Role of Neurokinin-1 Receptors in the Amygdala Author links open overlay panel
- Neuronal mitochondria modulation of LPS-induced neuroinflammation Micah Harland, Sandy Torres, Jingyi Liu and Xinglong Wang Journal of Neuroscience 14 January 2020, 2324-19; DOI: https://doi.org/10.1523/JNEUROSCI.2324-19.2020
5 of 8 citations matched to PubMed; remaining titles are listed without PMIDs.
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