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Based on an advanced Lentivirus vector RNA interference (RNAi) delivery system, our precisely algorithm-designed shRNA sequences achieve high-efficiency target gene silencing. This system is suitable for various cell models (including difficult-to-transfect primary cells and suspension cells) and in vivo animal experiments, providing reliable technical support for gene function research, disease mechanism analysis, and drug target validation.
Creative Biogene provides researchers with complete gene silencing solutions. From target selection to functional validation, each step is executed by an experienced professional team, ensuring the reliability and reproducibility of research results.
Figure 1. Scheme of the transfer and packaging vectors used to produce the third-generation lentivirus.
Precise shRNA Design Strategy
Our shRNA design combines multi-level screening algorithms with experimental validation systems:
Efficient Lentiviral Packaging System
Broad Application Compatibility
1Target Gene Analysis: Analyze client-provided gene information or perform a comprehensive target gene assessment for optimal knockdown regions.
2shRNA Design & Selection: Design 3-5 candidate shRNA sequences per target gene using our multi-algorithm optimization system and off-target analysis.
3Vector Construction: Clone selected shRNA sequences into the lentiviral backbone with appropriate promoters and selection markers.
4Quality Control Verification: Perform sequencing verification to confirm correct vector construction and provide detailed vector maps.
5Viral Packaging: Package lentivirus using our optimized four-plasmid co-transfection system in HEK293T cells.
6Purification & Concentration: Harvest viral supernatant and process through ultracentrifugation or tangential flow filtration for high-titer preparations.
7Functional Testing: Validate virus quality through titer determination, safety testing (mycoplasma/endotoxin), and optional silencing efficiency verification.
8Comprehensive Delivery: Provide final products including high-titer virus solution, control virus, detailed documentation, and personalized technical support.
Transform your gene silencing research with Creative Biogene's expert shRNA lentiviral solutions. Our specialized team of molecular biologists and viral vector experts is ready to guide you through every step of your gene knockdown project.
Q: How do you ensure shRNA targeting specificity and efficiency?
A: We employ a multi-level screening strategy to ensure shRNA's high specificity and efficiency: First, we predict candidate targets through bioinformatics algorithms, avoiding highly conserved regions and known functional domains; second, we design multiple candidate sequences for functional validation to screen the optimal sequence; finally, we optimize shRNA thermodynamic stability and processing efficiency by strictly controlling GC content, avoiding internal hairpin structures and repetitive sequences. Additionally, we provide scramble controls to verify specificity and confirm ≥70% silencing efficiency through qPCR/Western Blot.
Q: What advantages does the lentiviral system have compared to other gene delivery methods?
A: The lentiviral system has the following unique advantages:
(1) efficient transduction of both dividing and non-dividing cells, particularly suitable for difficult-to-transfect cells like primary cells and neurons;
(2) genomic integration capability, supporting long-term stable expression;
(3) relatively large vector capacity, suitable for complex expression cassettes;
(4) lower immunogenicity, reducing host immune responses;
(5) tissue targeting capability through pseudotype envelope modifications.
In comparison, lipid transfection efficiency depends on cell type and provides temporary expression, while AAV has higher safety but smaller vector capacity.
Q: How to choose the appropriate lentiviral vector backbone and reporter markers?
A: Vector selection should consider the following factors:
(1) Experimental purpose: non-integrating vectors for short-term experiments, integrating vectors for long-term experiments;
(2) Cell type: selecting appropriate promoters based on cell characteristics;
(3) Reporting requirements: fluorescent proteins (GFP/RFP) for direct observation, resistance markers (Puro/Hyg) for stable cell line screening;
(4) Expression level requirements: strong promoters (CMV) for overexpression experiments, endogenous promoters for physiological level expression. We can provide personalized recommendations based on your specific experimental needs.
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