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| Home -> Products & Services -> Custom Plasmid Vector Construction Services -> Custom Construction Service of AAV Expression Plasmid Vector | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Custom Construction Service of AAV Expression Plasmid Vector Adeno-associated virus (AAV) is a small, non-enveloped, single-stranded DNA virus belonging to the Parvoviridae family. Wild-type AAV is non-pathogenic and requires helper viruses (e.g., adenovirus or herpesvirus) for replication, ensuring inherent safety. Owing to its unique biological properties, AAV has become a star tool in gene therapy and basic research as a gene delivery vector. RGBiotech offers diverse serotypes AAV plasmid vectors and support customized tissue-targeted vector development. Consult us now at admin@rgbiotech.com to obtain a personalized AAV vector construction solution and boost your scientific research and clinical transformation! Why Choose RGBiotech? 1) Extensive Backbone Library: Plasmid vectors with diverse promoters (tissue-specific/inducible), reporter genes, and tags.2) Broad Serotype Coverage: AAV1-AAV13 and chimeric serotypes for precise tissue targeting. 4) Rigorous Quality & Control: Sequencing validation 5) Custom Solutions: End-to-end services from vector design to construction. 6) Fast Turnaround Read More Adeno-associated Virus (AAV) is a single-stranded DNA virus with natural replication deficiency. It requires helper viruses (such as adenovirus or herpesvirus) to complete replication. Recombinant AAV (rAAV) is genetically engineered to remove pathogenic genes while retaining the Inverted Terminal Repeats (ITRs), making it a safe and efficient gene delivery tool. 1. Features of AAV 1) High Safety: Wild-type AAV is non-pathogenic, and engineered vectors lack replication capability.2) Low Immunogenicity: Minimized host immune response compared to adenovirus or lentivirus. 3) Non-integrating: AAV genomes persist as episomal forms, eliminating insertional mutagenesis risks. 4) Persistent Expression: It exists as an episome in the cell nucleus and can sustain expression for more than 6 months in non-dividing cells. 5) Strong Tissue Tropism: There are 12 serotypes and more than 100 variants, which can precisely infect different tissues. 6) Excellent Diffusibility: Small in size, high in titer, and able to penetrate the blood-brain barrier, making it suitable for neuroscience research. 7) Vector Capacity: Supports target fragments ≤ 4.7 kb (including elements such as promoters). 8) Flexible Design: Can carry CDS regions, RNAi sequences, CRISPR tools, etc. 9) Multi-scenario Adaptability: Local injection (such as intravitreal, brain region) or systemic delivery (intravenous/intraperitoneal injection). 10 Precise Targeting: Tissue-specific delivery via serotype selection or promoter engineering. 2. AAV Serotypes & Tissue Tropism AAV has multiple serotypes (e.g., AAV1, AAV2, AAV5, AAV8, AAV9), each exhibiting distinct tissue and cell infection efficiencies.
If you are not sure about the optimal AAV serotype for your target cell or tissue, you could try using a AAV Rainbow Kit (containing a combination of multiple serotypes of AAV expressing different reporter genes) to optimize the injection protocol in the preliminary experiment. 3.Applications of AAV 1) Gene Therapy: For single-gene genetic diseases (such as hemophilia, spinal muscular atrophy), tumors, and metabolic diseases.2) Neuroscience Research: Delivery of optogenetic, chemogenetic, and calcium indicators to analyze the functions of neural circuits. 3) Animal Model Construction: Achieve gene overexpression or silencing through tissue-specific promoters. 4) Drug Screening: Efficiently infect primary cells or organoids, accelerating drug development. 4. Elements of AAV Plasmid Expression Vector 1) Inverted Terminal Repeats (ITRs): Mediate virus replication and packaging.2) Promoters: Broad-spectrum (such as CMV, EF1α) or tissue-specific (such as hSyn, Alb). 3) Target Gene: Coding region (CDS) or functional sequence (such as sgRNA, shRNA). 4) Reporter Gene: Fluorescent proteins (GFP, mCherry). 5. AAV Virus Packaging Process 1) Plasmid Construction: Clone the target gene into the AAV expression or transfer vector, together with the helper plasmid (Rep/Cap) and the packaging plasmid.2) Cell Transfection: Co-transfect HEK293 cells with the AAV expression plasmid, the rep/cap plasmid and the helper plasmid to initiate virus assembly. 3) Virus Harvesting: Collect the cell lysate and supernatant 48-72 hours later. 4) Purification and Concentration: Use CsCl gradient centrifugation, affinity chromatography, or ultrafiltration technology to remove impurities and purify viral particles. 5) Quality Control: Assess titer (genome copies/mL, GC/mL) by qPCR, purity (host protein/DNA residues) by SDS-PAGE and assay kits, endotoxin (LAL), potency (in vitro infection assays). |
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