Pseudotyping Services of Lentiviral Vectors with Rabies Virus
Pseudotyping as a cornerstone technique in modern virology and genetic engineering, enables the structural adjustment of viral vectors according to specific research and treatment requirements1,2. By replacing the natural envelope protein of lentiviral vectors with the rabies virus glycoprotein (RV-G), researchers can utilize an efficient retrograde transport system to achieve a unique function, allowing these vectors to move retrogradely along the axons of neurons. Therefore, rabies virus phage-displayed lentiviral vectors (RV-LVs) have become an indispensable tool for mapping complex neural circuit diagrams, delivering genetic material to specific neuronal populations, and developing novel gene therapies for neurological diseases.
Fig.1 Applying pseudotyped LVs for cancer gene therapy 1,2
At Creative Biolabs, we focus on the design, production and validation of high-quality rabies pseudotyped lentiviral vectors. Our comprehensive service platform combines the most advanced molecular virology with strict quality control, providing researchers with reliable and directly usable viral vectors that maintain stable performance in both in vitro and in vivo environments. Whether you are studying basic neurobiological mechanisms or developing next-generation therapeutic strategies, we can offer comprehensive design solutions to help you solve problems quickly and efficiently. Fig.2 shows the design process about pseudotyping of lentiviral vectors with rabies virus glycoprotein for neural circuit research and gene therapy.
Fig.2 Pseudotyping of lentiviral vectors with rabies virus glycoprotein for neural circuit research and gene therapy (Creative Biolabs Original)
Our Workflow: From Concept to Ready-to-Use Vectors
We follow a structured, phase-gated process to ensure transparency, quality, and reproducibility at every stage of vector development:
1. Consultation & Vector Design
When initiating each project, we will thoroughly discuss your research objectives, experimental system and specific requirements with you. Our experts will provide you with the best guidance on vector design, including promoter selection, reporter gene selection and cassette configuration, to maximize your experimental results.
2. Plasmid Construction & Validation
Our molecular biology team will employ advanced cloning techniques to clone your gene fragment into an optimized lentiviral backbone vector. All the constructed entities have been fully verified by Sanger sequencing to ensure the complete accuracy and genetic integrity of the sequences.
3. Virus Production & Purification
We employ an expandable production system, using HEK293T cells and advanced transfection reagents to prepare high-titer virus samples. Subsequently, the samples are purified through a benzyl alcohol gradient ultracentrifugation to remove impurities and enrich fully functional virus particles.
4. Comprehensive Quality Control
- Quantitative PCR for genomic titer determination
- p24 ELISA for physical particle quantification
- Functional titration in relevant cell lines
- Endotoxin and sterility testing
- SDS-PAGE analysis for protein purity assessment
5. Delivery & Documentation
After you complete the order, we will immediately send your virus samples according to the concentration you specified. These samples will be safely packaged with dry ice to ensure their stability. Each delivery will come with a detailed analysis certificate, which includes complete quality control results, as well as a comprehensive project report that outlines the entire production process and various characteristic data.
Key Advantages of Our Pseudotyping Services
- Superior Retrograde Transport Efficiency: Our RV-LVs demonstrate robust retrograde transport, efficiently transducing neurons that project to the injection site—making them ideal for detailed connectome mapping and functional analysis of neural circuits.
- High Transduction Efficiency and Expression Levels: Through optimized production protocols and rigorous QC, we ensure that our vectors deliver robust gene expression in both in vitro and in vivo applications. Our vectors consistently achieve high transduction rates across various neuronal cell types.
- Rigorous Quality Assurance: We implement a multi-tiered QC approach that exceeds industry standards. Every viral batch is thoroughly characterized for titer, purity, functionality, and safety, providing you with reliable, reproducible results across experiments.
- Expert Scientific Collaboration: Our team includes experienced virologists and neuroscientists who provide end-to-end project support from experimental design to data interpretation. We function as an extension of your research team, offering technical insights and troubleshooting assistance throughout your project.
- Flexible and Customizable Solutions: We understand that every research project has unique requirements. Our services are highly adaptable to accommodate specific promoter choices, reporter combinations, serotype preferences, and purification specifications.
Gene Therapy Development
The retrograde transport capability of RV-LVs makes them ideal vehicles for delivering therapeutic genes to specific neuronal populations affected by neurological disorders.
1. Applications include:
- Targeted delivery of neuroprotective factors
- Selective modulation of disease-related circuitry
- Development of novel treatments for Parkinson's disease, epilepsy, and pain disorders
- Functional Genomics and Optogenetics
2. RV-LVs enable precise manipulation and monitoring of neural activity in defined circuits:
- Expression of optogenetic tools for photostimulation
- Delivery of chemogenetic receptors for remote neural control
- Introduction of calcium indicators for neural activity imaging
- CRISPR-based functional screening in specific neuronal populations
- Behavioral Neuroscience
3. These vectors facilitate circuit-level interrogation of behavioral mechanisms:
- Dissecting neural circuits underlying learning and memory
- Investigating reward and motivation pathways
- Studying neural substrates of affective disorders
- Examining sensorimotor processing circuits
Client Testimonials
Frequently Asked Questions
Q: What are the specific advantages of using Rabies-G pseudotyped lentivirus compared to other envelopes?
A: Rabies-G confers efficient retrograde trafficking capabilities, enabling transduction of neurons that project to the injection site—a property absent in most other pseudotypes. This makes RV-LVs particularly valuable for neural circuit mapping studies. Additionally, Rabies-G pseudotyped vectors exhibit excellent stability and broad neuronal tropism, making them suitable for targeting diverse neuronal populations.
Q: Can these vectors be used in non-neuronal tissues or applications outside neuroscience?
A: While Rabies-G is particularly renowned for its neuronal tropism and retrograde transport properties, we also offer pseudotyping services with other envelope proteins (e.g., VSV-G, MLV, Ross River Virus) for applications requiring broad tropism or targeting specific non-neuronal cell types. Our team can advise on the optimal pseudotype for your specific research needs.
Q: Do you offer single-cycle (G-deleted) Rabies systems for restricted spread?
A: Yes, we provide both full-length and G-deleted Rabies glycoprotein systems. The G-deleted system offers enhanced biosafety and restricted spread, limiting transduction to directly infected cells without secondary transmission. This is particularly valuable for precise circuit mapping and reduced off-target effects.
Q: What level of documentation and quality control data is provided with the vectors?
A: Each viral preparation is accompanied by a comprehensive Certificate of Analysis that includes detailed information on genomic and physical titers, endotoxin levels, sterility testing results, and functional validation data. Additionally, we provide a complete project report documenting all production and QC processes, ensuring full transparency and reproducibility.
Q: How do I determine the appropriate titer and volume for my experiments?
A: Our technical team provides personalized guidance on dosing based on your specific experimental system, target region, and application. We consider factors such as injection volume, neuronal density, and expression level requirements to recommend optimal vector quantities for your study.
Supported by Scientific Excellence
The application of Rabies pseudotyped vectors represents a cutting-edge methodology in neuroscience research. Recent studies have demonstrated their utility in mesoscale connectomics and functional interrogation of neural pathways. Our vectors are developed in alignment with these advanced methodologies, incorporating the latest insights from virology and neuroscience to ensure optimal performance. We maintain an active research and development program to continuously improve our vector systems and stay at the forefront of technological advancements.
Begin Your Research Journey With Us
Unlock the full potential of neural circuit research with custom pseudotyped lentiviral vectors from Creative Biolabs. Our comprehensive service platform combines scientific expertise with rigorous quality control to deliver viral vectors that meet the most demanding research standards. Whether you're launching a new research direction or scaling up existing studies, our team is ready to collaborate with you to design, produce, and validate vectors that will drive your discoveries forward.
Contact us today to schedule a confidential consultation with our technical team. Let us help you accelerate your research timeline and achieve your scientific goals with precision-engineered viral vector solutions.
References
- Deng,Longfei, et al. "Pseudotyped lentiviral vectors: Ready for translation into targeted cancer gene therapy?" Genes & Diseases (2023) 10, 1937-1955. https://doi.org/10.1016/j.gendis.2022.03.007
- Distributed under Open Access license CC BY 4.0, without modification.
