| Field | Specification |
|---|---|
| Form | Liquid |
| Function | |
| Plasmid Backbone | |
| Product Type | |
| Production System | |
| Promoter | |
| Reporter | |
| Storage |
Overview
AAV-GFP (AAV serotype 2) AAV (AAV2-GFP) is an AAV vector packaged in AAV2 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): AAV2. efficient neuronal transduction with limited spread; widely used for in vitro work and for focal CNS injections.
- 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 AAV2?
What controls should I include alongside this AAV?
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Selected References
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- Convection-enhanced delivery of AAV2 in white matter¿A novel method for gene delivery to cerebral cortex
- Smith LN, Jedynak JP, Fontenot MR, et al. Fragile X mental retardation protein regulates synaptic and behavioral plasticity to repeated cocaine administration. Neuron 2014. PMID: 24811383
- Erdos B, Backes I, McCowan ML, et al. Brain-derived neurotrophic factor modulates angiotensin signaling in the hypothalamus to increase blood pressure in rats. Am J Physiol Heart Circ Physiol 2015. PMID: 25576628
- Huntingto’s disease: Neural dysfunction linked to inositol polyphosphate multikinase
- Toda C, Kim JD, Impellizzeri D, et al. UCP2 Regulates Mitochondrial Fission and Ventromedial Nucleus Control of Glucose Responsiveness. Cell 2016. PMID: 26919426
- Lee AS, De Jesús-Cortés H, Kabir ZD, et al. The Neuropsychiatric Disease-Associated Gene cacna1c Mediates Survival of Young Hippocampal Neurons. eNeuro 2016. PMID: 27066530
- MacKenzie G, O'Toole KK, Moss SJ, et al. Compromised GABAergic inhibition contributes to tumor-associated epilepsy. Epilepsy Res 2016. PMID: 27513374
- Reassembly of Excitable Domains after CNS Axon Regeneration. PMID: 27581456
- Wang Y, Li WY, Jia H, et al. KLF7-transfected Schwann cell graft transplantation promotes sciatic nerve regeneration. Neuroscience 2017. PMID: 27826105
- Lewis JE, Brameld JM, Hill P, et al. Hypothalamic over-expression of VGF in the Siberian hamster increases energy expenditure and reduces body weight gain. PLoS One 2017. PMID: 28235047
- Kim JD, Toda C, Ramírez CM, et al. Hypothalamic Ventromedial Lin28a Enhances Glucose Metabolism in Diet-Induced Obesity. Diabetes 2017. PMID: 28550108
- Melón LC, Hooper A, Yang X, et al. Inability to suppress the stress-induced activation of the HPA axis during the peripartum period engenders deficits in postpartum behaviors in mice. Psychoneuroendocrinology 2018. PMID: 29274662
- Bruschetta G, Kim JD, Diano S, et al. Overexpression of melanocortin 2 receptor accessory protein 2 (MRAP2) in adult paraventricular MC4R neurons regulates energy intake and expenditure. Mol Metab 2018. PMID: 30352741
- Darnieder LM, Melón LC, Do T, et al. Female-specific decreases in alcohol binge-like drinking resulting from GABA(A) receptor delta-subunit knockdown in the VTA. Sci Rep 2019. PMID: 31147611
- Cai WT, Yoon HS, Lee S, et al. Repeated exposure to methiopropamine increases dendritic spine density in the rat nucleus accumbens core. Neurochem Int 2019. PMID: 31176680
- Deficiency in BDNF/TrkB Neurotrophic Activity Stimulates d-Secretase by Upregulating C/EBPß in Alzheimer’s Disease
- AAV8-vectored suprachoroidal gene transfer produces widespread ocular transgene expression. PMID: 31408444
- HDAC inhibition leads to age-dependent opposite regenerative effect upon PTEN deletion in rubrospinal axons after SCI.
- Lombaert IMA, Patel VN, Jones CE, et al. CERE-120 Prevents Irradiation-Induced Hypofunction and Restores Immune Homeostasis in Porcine Salivary Glands. Mol Ther Methods Clin Dev 2020. PMID: 32953934
- Bruschetta G, Jin S, Liu ZW, et al. MC(4)R Signaling in Dorsal Raphe Nucleus Controls Feeding, Anxiety, and Depression. Cell Rep 2020. PMID: 33053350
- Umanah GKE, Ghasemi M, Yin X, et al. AMPA Receptor Surface Expression Is Regulated by S-Nitrosylation of Thorase and Transnitrosylation of NSF. Cell Rep 2020. PMID: 33147468
19 of 23 citations matched to PubMed; remaining titles are listed without PMIDs.
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