rAAV-TRE3g
The TRE3g promoter (tetracycline-responsive element version 3, enhanced) is a component of the Tet-On/Tet-Off system, which enables inducible and tightly controlled gene expression. Recombinant adeno-associated viruses (rAAVs) incorporating TRE3g promoters allow for doxycycline-dependent transgene regulation, providing precise temporal control over gene expression.
Features of rAAV-TRE3g
- Inducible Gene Expression:
- TRE3g requires the presence of the rtTA (reverse tetracycline transactivator) protein and doxycycline to initiate transcription. Expression ceases when doxycycline is removed.
- Tight on/off control ensures minimal leaky expression in the absence of doxycycline.
- Enhanced Sensitivity:
- TRE3g is an improved version of earlier TRE promoters, offering reduced background expression and higher responsiveness to doxycycline.
- Temporal Control:
- Enables precise timing of transgene expression, critical for applications where constitutive expression is undesirable.
- Compatibility with Diverse Systems:
- Can be combined with neuron-, glia-, or tissue-specific promoters driving rtTA expression for cell-type-specific and inducible transgene control.
Applications of rAAV-TRE3g
- Neuroscience Research:
- Circuit Manipulation: Inducible delivery of optogenetic or chemogenetic tools (e.g., channelrhodopsins, DREADDs) to specific neural circuits.
- Calcium Imaging: Temporal control over calcium indicators (e.g., GCaMP) for activity-dependent studies.
- Gene Therapy:
- Therapeutic Regulation: Controlled delivery of therapeutic genes, allowing regulation of dosage and timing in diseases like epilepsy or neurodegeneration.
- Safety: Reduces risks associated with constitutive expression of potent transgenes by enabling doxycycline-dependent expression.
- Disease Modeling:
- Inducible models of disease that require temporally regulated gene expression for specific developmental stages or experimental conditions.
- CRISPR/Cas9 Genome Editing:
- Controlled activation of CRISPR/Cas9 systems to minimize off-target effects or study time-sensitive genetic interventions.
rAAV-TRE3g is an advanced tool for precise, inducible gene expression, offering robust control over timing and dosage. It is especially useful in applications requiring temporal regulation of gene expression, such as functional studies, disease modeling, and therapeutic development. When combined with tissue-specific rtTA expression, it provides unparalleled flexibility in gene delivery and regulation.
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