Overview of rAAV-CAG

Recombinant Adeno-Associated Virus (rAAV) vectors are widely utilized in gene therapy and neuroscience research due to their ability to efficiently deliver genetic material into host cells. One of the key elements determining the success of these vectors is the promoter used to drive gene expression. The rAAV-CAG vector utilizes the CAG promoter, a hybrid promoter derived from the Cytomegalovirus immediate-early enhancer, the chicken β-actin promoter, and the rabbit β-globin splice acceptor.

Technical Content on rAAV-CAG

Vector Construction:

• The rAAV-CAG vector is engineered to include the CAG promoter upstream of the gene of interest.
• The CAG promoter combines elements to ensure high levels of transgene expression in a wide range of cell types, including neurons.
• This promoter enhances the transcriptional activity, making it suitable for studies requiring robust gene expression.

Mechanism of Action:

• Upon delivery into the host cell, the rAAV vector utilizes the cellular machinery for the synthesis of the transgene.
• The CAG promoter's strong enhancer elements facilitate the recruitment of transcription factors, promoting efficient transcription initiation.
• The β-globin splice acceptor ensures proper mRNA processing, enhancing the stability and translation of the transcript.

Applications:

• Gene Therapy: rAAV-CAG vectors are extensively used for delivering therapeutic genes in models of genetic disorders. The high expression levels driven by the CAG promoter are advantageous for diseases requiring substantial gene product to achieve therapeutic effects.
• Neuroscience Research: In neuroscience, rAAV-CAG is employed to express genes in neuronal populations, aiding in studies of neural function, behavior, and disease mechanisms. The promoter's efficiency in neurons makes it particularly valuable for these applications.
• Functional Studies: Researchers utilize rAAV-CAG to overexpress proteins, investigate gene function, and model diseases. Its ability to drive high expression levels is crucial for experiments where the phenotype is dose-dependent.

Advantages:

• Broad Tropism: The CAG promoter is active in various cell types, including neurons, muscle cells, and hepatocytes, making the vector versatile for different research applications.
• High Expression Levels: The hybrid nature of the CAG promoter ensures robust and sustained transgene expression, which is essential for both in vitro and in vivo studies.
• Compatibility with AAV Serotypes: The rAAV-CAG vector can be packaged with different AAV serotypes, each having distinct tissue tropisms, allowing tailored delivery based on the target tissue.