AAV Vector Design Service for RNAi Delivery

Introduction

To address challenges in therapeutic gene delivery, long-term expression, and gene dosage control, our AAV Vectors for Delivery of RNAi service provides a safe, efficient platform for targeted gene silencing. Using advanced vector engineering and RNAi technology, it enables precise control of gene expression, laying the groundwork for effective therapies for genetic diseases and cancer. We also offer a clear, reliable pathway from concept to functional AAV vector, with specific solutions and support at every step.

AAV Vectors for Delivery of RNAi

Core Components of RNAi

RNAi (RNA interference) is a gene silencing mechanism mediated by small RNAs in eukaryotes. Its core components can be divided into two parts: key molecules and the mechanism of action.

Fig.1 The components that make up RNA interference (RNAi) include the biosynthesis of small RNAs, the formation of RISCs, and the effector stage.Distributed under CC BY-SA 4.0, from Wiki, without modification.

• Key Molecules
  • Small Interfering RNA (siRNA): Double-stranded RNA, usually produced by the processing of exogenous nucleic acids (e.g., viral RNA), with a length of approximately 21-23 nucleotides. It can accurately recognize complementary target mRNAs.
  • MicroRNA (miRNA): Derived from the processing of endogenous single-stranded RNA precursors (pre-miRNA), with a length of about 20-25 nucleotides. It can bind to target mRNAs through incomplete complementarity to regulate the expression of endogenous genes.
  • Core Protein Complex (RISC): Short for RNA-induced silencing complex, which contains proteins such as Argonaute (Ago). It can bind to siRNA/miRNA and mediate the degradation or translational inhibition of target mRNAs.
• Mechanism of Action

1. Step 1 (Processing): Exogenous double-stranded RNA is cleaved into siRNA by Dicer enzyme; endogenous pri-miRNA is sequentially processed into mature miRNA by Drosha and Dicer enzymes.
2. Step 2 (Assembly): siRNA/miRNA binds to Ago protein to form an active RISC.
3. Step 3 (Silencing): RISC binds to target mRNA through base complementarity. siRNA causes the degradation of target mRNA due to complete complementarity, while miRNA inhibits the translation of target mRNA due to incomplete complementarity. Ultimately, gene expression silencing is achieved.

How AAV-Mediated RNAi Delivery Works

• Vector Construction: Insert DNA sequences encoding siRNA, miRNA, or shRNA (processed into siRNA in cells) into the AAV genome; select AAV serotype matching target tissue (e.g., AAV8 for liver, AAV9 for CNS).
• In Vivo Delivery: Administer recombinant AAV (carrying RNAi elements) via intravenous/local injection (e.g., intracerebral, intramuscular); AAV infects target cells specifically via tissue tropism.
• Element Expression and Processing: AAV genome (with RNAi sequences) enters the target cell nucleus, exists stably as a non-integrating "episome" (high safety), and uses the host transcription system to synthesize shRNA/miRNA precursors.
• Initiation of RNAi Silencing: shRNA is cleaved into siRNA by Dicer, and assembles with Ago into RISC; miRNA precursors become mature miRNA and form RISC. Both silence target genes via mRNA degradation or translation inhibition, achieving disease treatment (e.g., inhibiting pathogenic gene expression).

Advantages

When combined, AAV and RNAi offer a powerful one-two punch. AAV ensures efficient, long-term delivery of the RNAi machinery to the target cells, and RNAi provides the specific knockdown of the desired gene. This approach offers sustained therapeutic effect from a single administration, which is a major advantage for treating chronic conditions.

Workflow

1. Required Starting Materials: To initiate your project, we require:
   • The gene sequence you wish to silence (target gene).
   • Any specific cell lines or animal models you plan to use for validation?
   • Your desired delivery route (e.g., systemic, local).
2. Project Consultation & Design: Team reviews project goals/materials to design optimal RNAi strategy, selecting shRNA/miRNA sequences, promoters, and AAV serotype matching target tissue.
3. Vector Construction: Synthesizes a designed genetic cassette, clones it into a high-quality AAV plasmid (with elements for high RNAi trigger expression).
4. Virus Production & Purification: Generates high-titer research-grade AAV via an advanced platform, purifying to remove impurities and empty capsids.
5. Quality Control & Validation: Conducts rigorous QC (viral titer measurement, payload integrity confirmation) and in vitro validation for efficient gene silencing.
6. Final Delivery: Provides validated AAV vector plus detailed report (vector sequence, titer, QC results).
7. Final Deliverables: Clients receive purified, ready-to-use AAV, a comprehensive project report, and in vitro/in vivo application protocols.
8. Estimated Timeframe: 8–12 weeks (varies by project complexity/design needs); precise timeline available in initial consultation.

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What We Can Offer

At Creative Biolabs, we recognize that every gene therapy project is unique. Our services go beyond standard vector production to provide you with a truly customized and comprehensive solution for your RNAi delivery needs. We collaborate with you from the initial design phase to final delivery, ensuring that your research goals are met with precision and efficiency.

Our Advantage

Customer Reviews

"Using Creative Biolabs' AAV RNAi service in our research has significantly improved the efficiency of our gene knockdown studies. The high-titer vectors and their rigorous QC facilitated our in vivo work, allowing us to achieve stable gene silencing in our animal models. Their documentation was thorough and their support was excellent."

Dr. Miller, Leading Researcher at a Top University.

"We were facing challenges with off-target effects using alternative methods. The targeted AAV vectors from Creative Biolabs provided the precision we needed. The clear advantage was the ability to modulate gene expression in a tissue-specific manner, which was critical for our work on a complex genetic disease. The final product and accompanying data were outstanding."

Dr. Carter, Senior Scientist at a Biotech Firm.

"The robust and reproducible results from Creative Biolabs' AAV vectors were a game-changer for our cancer research. We needed to specifically silence a gene in tumors without affecting healthy tissue, and the vectors delivered a dose-dependent knockdown, similar to published data. Their expertise and high-quality materials saved us months of optimization."

Dr. Chen, Principal Investigator.

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FAQs

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