Recombinant adeno-associated virus (rAAV) vectors are crucial tools in both neuroscience and gene therapy research due to their versatility, safety, and efficiency. Here, we present ten of the most popular rAAV vectors, detailing their structures and explaining why they are widely used in the scientific community.
rAAV-CMV-GFP-WPRE-bGH polyA
- Promoter: CMV (Cytomegalovirus)
- Reporter: GFP (Green Fluorescent Protein)
- Enhancer: WPRE (Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element)
- PolyA: bGH (bovine Growth Hormone polyadenylation signal)
rAAV-Syn-GFP-WPRE-bGH polyA
- Promoter: Syn (Synapsin)
- Reporter: GFP
- Enhancer: WPRE
- PolyA: bGH
rAAV-hSyn-ChR2(H134R)-EYFP-WPRE-bGH polyA
- Promoter: hSyn (human Synapsin)
- Reporter: ChR2(H134R)-EYFP (Channelrhodopsin-2 H134R variant fused with Enhanced Yellow Fluorescent Protein)
- Enhancer: WPRE
- PolyA: bGH
rAAV-CAG-tdTomato-WPRE-bGH polyA
- Promoter: CAG (CMV early enhancer/chicken β-actin)
- Reporter: tdTomato (Tandem dimer Tomato Fluorescent Protein)
- Enhancer: WPRE
- PolyA: bGH
rAAV-CMV-mCherry-WPRE-bGH polyA
- Promoter: CMV
- Reporter: mCherry (Monomeric Cherry Fluorescent Protein)
- Enhancer: WPRE
- PolyA: bGH
rAAV-hSyn-hChR2(H134R)-mCherry-WPRE-bGH polyA
- Promoter: hSyn
- Reporter: hChR2(H134R)-mCherry (Human Channelrhodopsin-2 H134R variant fused with mCherry)
- Enhancer: WPRE
- PolyA: bGH
rAAV-hSyn-DIO-GCaMP6f-WPRE-bGH polyA
- Promoter: hSyn
- Reporter: DIO-GCaMP6f (Cre-dependent GCaMP6f calcium indicator)
- Enhancer: WPRE
- PolyA: bGH
rAAV-CaMKII-GFP-WPRE-bGH polyA
- Promoter: CaMKII (Calcium/Calmodulin-dependent protein kinase II)
- Reporter: GFP
- Enhancer: WPRE
- PolyA: bGH
rAAV-hSyn-ArchT-GFP-WPRE-bGH polyA
- Promoter: hSyn
- Reporter: ArchT-GFP (Archaerhodopsin-T fused with GFP)
- Enhancer: WPRE
- PolyA: bGH
rAAV-hSyn-DIO-mCherry-WPRE-bGH polyA
- Promoter: hSyn
- Reporter: DIO-mCherry (Cre-dependent mCherry)
- Enhancer: WPRE
- PolyA: bGH
Why These rAAV Vectors Are Popular
These rAAV vectors are favored for several reasons related to their design and functionality:
- Promoters:
- CMV: Known for driving high levels of gene expression across a variety of cell types.
- hSyn: Targets expression specifically to neurons, essential for neuroscience research.
- CAG: Promotes high expression levels in a broad range of tissues.
- CaMKII: Specific to excitatory neurons in the central nervous system, allowing for targeted expression.
- Reporters:
- GFP, EYFP, mCherry, and tdTomato: Fluorescent proteins widely used for their strong fluorescence and ease of detection.
- ChR2 and ArchT: Opsins crucial for optogenetics, enabling control of neuronal activity with light.
- GCaMP6f: A calcium indicator used for imaging neuronal activity.
- Enhancers:
- WPRE: Enhances the stability and translation of mRNA, leading to higher expression levels of the transgene.
- Polyadenylation signals:
- bGH polyA: Ensures efficient termination and polyadenylation of mRNA, which is crucial for stability and translation efficiency.
- Versatility and Specificity:
- The combination of different promoters and reporters allows for precise control over gene expression. For example, hSyn promotes neuron-specific expression, while CMV and CAG promote broad tissue expression.
- Established Efficacy:
- These vectors have been validated in numerous studies, demonstrating reliable and reproducible results, making them preferred choices for researchers.
- Community and Tool Availability:
- Extensive documentation, availability of plasmids from repositories like AffiGEN, and a large body of literature using these vectors make them accessible and easier to troubleshoot.
- Safety and Efficiency:
- rAAV vectors are known for their safety profile, low immunogenicity, and ability to transduce both dividing and non-dividing cells, making them suitable for in vivo applications.
In conclusion, the combination of strong promoters, versatile reporters, effective enhancers, and reliable polyadenylation signals makes these rAAV vectors essential tools for a wide range of applications in research and therapeutic settings.