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Creative Biogene provides a wide selection of recombinase AAV particles containing key genome engineering proteins including Cas9, Cpf1, Cre, and FLPo. These optimized nuclease and recombinase AAVs enable precise manipulation of cell genomes through targeted DNA cleavage and recombination events. Key advantages offered by Creative Biogene's reagents include validated activity in mammalian cells, robust viral production using triple-plasmid transfection, and custom recombinant AAV services. Tight regulation of recombinase expression can be achieved using tissue-specific or inducible promoters. The recombinase AAVs represent powerful tools for diverse applications from gene function studies to gene therapy.
Site-specific nucleases like Cas9 and Cpf1 can be introduced into cells via AAV vectors to achieve genome editing at desired loci specified by the guide RNA sequence. This allows targeted gene knockout, mutation, or correction. The principle involves cloning the nuclease open reading frames into AAV transfer plasmids and subsequent transduction. Likewise, recombinases like Cre and FLPo recognize short sequences (loxP or FRT sites) flanking genomic regions of interest. AAV delivery of these recombinases to cells containing pre-engineered loxP or FRT sites enables excision, inversion, or translocation of the intervening sequence. Tissue-specific Cre-expression allows cell-specific recombination. Overall, Creative Biogene's recombinase AAVs provide robust tools for genetic manipulation, including gene function studies, generating animal models, and therapeutic correction of disease-causing mutations.
You can use our recombinant AAV particles to conveniently complete the following tasks:
Case Study 1
Alterations in social behavior are core symptoms of major developmental neuropsychiatric diseases such as autism spectrum disorders or schizophrenia. Serum response factor (SRF) is a major transcription factor in the brain. The researchers inserted a CreERT2 fusion construct at the ATG of the CaMKIIα gene to regulate Cre recombinase expression under the control of the endogenous CaMKIIα gene. Neurons expressing Cre recombinase exhibited increased spine length, a reduced percentage of mature dendritic spines, and an augmented presence of immature protrusions, mirroring the outcomes observed with shRNA. This independent validation confirms the role of the Serum Response Factor (SRF) in promoting the maturation of structural dendritic spines. Significantly divergent dendritic spine densities were observed among the analyzed groups in the experiment.
Figure 1. The research confirmed that AAV-shSRF efficiently reduces SRF in hippocampal cultures. The constructs AAV-shSRF and AAV-Cre demonstrated effective reduction of SRF expression in vitro neuronal settings. (Roszkowska M, et al., 2021).
Case Study 2
Astrocytes play an important role in increasing synaptic plasticity, regulating endogenous homeostasis, and contributing to neuroprotection but become overactivated or apoptotic in persistent neuroinflammatory responses or pathological conditions. The researchers made mice to induce astrocyte apoptosis using AAV-Cre. Localized cell loss in the hippocampus caused transient deficits and reactive gliosis that recovered within 2 weeks. It provides a model to study reactive astrocyte regeneration.
Figure 2. The researchers generated a transgenic caspase-3 mouse using a Cre-inducible vector. They validated astrocyte apoptosis induction in the mice using PCR, immunochemistry, and Western blotting. (Kim S C, et al., 2022).
Case study 3
Accumulation of excess nutrients hampers proper liver function and is linked to nonalcoholic fatty liver disease (NAFLD) in obesity.The researchers found SUMOylation regulates fasting metabolism via Prox1 in the liver. An obesity model showed defective Prox1 SUMOylation, while a SUMO-defective mutant maintained fasting pathways. Prox1 SUMOylation may be a nutrient-sensing switch.
Figure 3. The researchers injected a Prox1 SUMOylation mouse model with AAV-Cre, showing abolished Prox1 SUMOylation versus controls. This verifies the model for studying Prox1 SUMOylation in liver metabolism. (Alfaro A J, et al., 2023).
Q: What are Recombinase AAV particles?
A: Recombinase AAV particles contain the gene for a site-specific recombinase enzyme such as Cre or Flp. When delivered to cells, the recombinase catalyzes recombination between specific DNA sequences to induce effects such as gene deletion, inversion or chromosomal translocation. This allows spatial and temporal control of gene editing events by delivery of recombinase AAVs.
Q: What is the process of rAAV transduction of cells?
A: (1) Binding to a primary receptor (e.g. heparan sulfate proteoglycan) and co-receptors on the cell surface.
(2) Endocytosis of the AAV particle via clathrin-coated pits or other mechanisms.
(3) Trafficking of the endosome to the perinuclear region.
(4) Endosomal escape of the AAV capsid to the cytoplasm.
(5) Translocation of the AAV to the nucleus.
(6) Uncoating of the capsid to release the viral genome.
(7) Conversion of the single-stranded AAV genome into double-stranded DNA.
(8) Transcription and expression of the transgene.
Q: What are the advantages of rAAV vectors?
A: (1) Low immunogenicity and toxicity
(2) Ability to transduce both dividing and non-dividing cells
(3) Long-term transgene expression
(4) Minimal risk of insertional mutagenesis
(5) Capacity to package up to ~4.5 kb transgene constructs
(6) Availability of many natural and engineered capsid serotypes
(7) Lack of associated human disease
Q: How is the rAAV serotype selected?
A: (1) Natural tropism of the capsid for the desired cells/tissue
(2) Pre-existing immunity against different capsids in the host
(3) Performance in preclinical models for the intended tissue
(4) Capsid-specific differences in transduction efficiency, kinetics, and immunogenicity
(5) Availability of specific capsid variants and engineered capsids
(6) Payload capacity needed for the transgene construct
(7) Testing different serotypes in vitro and in vivo is recommended to empirically determine the optimal capsid.
| Cat.No. | Product Name | Price |
|---|---|---|
| AAV00020Z | Cre Adeno-associated virus(AAV Serotype 5) | Inquiry |
| AAV00044Z | Cre Adeno-associated virus(AAV Serotype 1) | Inquiry |
| AAV00045Z | Cre Adeno-associated virus(AAV Serotype 2) | Inquiry |
| AAV00046Z | Cre Adeno-associated virus(AAV Serotype 6) | Inquiry |
| AAV00047Z | Cre Adeno-associated virus(AAV Serotype 8) | Inquiry |
| AAV00048Z | Cre Adeno-associated virus(AAV Serotype 9) | Inquiry |
| AAV00061Z | Cre-GFP Adeno-associated virus(AAV Serotype 1) | Inquiry |
| AAV00062Z | Cre-GFP Adeno-associated virus(AAV Serotype 2) | Inquiry |
| AAV00063Z | Cre-GFP Adeno-associated virus(AAV Serotype 5) | Inquiry |
| AAV00064Z | Cre-GFP Adeno-associated virus(AAV Serotype 6) | Inquiry |
| AAV00065Z | Cre-GFP Adeno-associated virus(AAV Serotype 8) | Inquiry |
| AAV00066Z | Cre-GFP Adeno-associated virus(AAV Serotype 9) | Inquiry |
| AAV00100Z | Synapsin-Cre-GFP AAV (Serotype 8) | Inquiry |
| AAV00111Z | AAV9-CMV-Cas9 | Inquiry |
| AAV00123Z | scAAV1-Cre | Inquiry |
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