Gene Kockdown Validation
Introduction
Gene silencing is an important regulation mechanism that reduce or suppress the expression of specific genes at the transcriptional or post-transcriptional levels. Transcriptional gene silencing is often triggered by DNA methylation, heterochromatin and position effect, and while Post-transcriptional gene silencing is triggered by antisense RNA or RNAi. RNAi has the two main advantages: the high degrees of efficiency and specificity, and shows great promise in studying gene function and drug discovery in the post genomic era. RNAi is a gene-silencing phenomenon whereby double-stranded RNA (dsRNA) triggers the sequence-specific degradation of homologous mRNA. In RNAi experiments, since the gene silencing efficiency of the siRNA is unpredictable, therefore, the choice of siRNA with high efficiency and specificity is critical to successful gene silencing.
Service Details
WZ Biosciences can provide personalized Gene Kockdown Validation Services for you according to your research needs, including Custom shRNA Validation and efficient shRNA Validation.
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Cat No.
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Type
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Validation Method
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Price
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Turnaround Time
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WZ090001
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Custom shRNA Validation
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Real-time PCR
Western blot
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$400/test
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1-2weeks
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WZ090002
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Efficient shRNA Validation
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Real-time PCR
Western blot
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$400/test
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1-2weeks
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*Interested in above Kockdown Validation Services? Please feel free to contact us to “Request a quote”.
Related services： shRNA Cloning, Adenovirus Packaging, Lentivirus Packaging, AAV Packaging,
What should I do if there is no suitable target cell for efficient shRNA validation?
When researchers validate the efficiency of the shRNAs targeting mouse(or rat) genes, they usually have no suitable target cells for efficient shRNA validation, what should be done? For the efficient siRNA screening, researchers typically test multiple shRNAs(3 to 4) per target gene. In order to validate the specificity and efficiency of shRNAs in vitro, we can co-transfect the shRNA expression plasmids and their corresponding cDNA expression plasmids into HEK293 cells. Validation of shRNA knockdown efficiency was assessed by conventional molecular biology methods.
Service Process
Related Resorces
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《Transfection Reagent Instructions》
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The Factors Affecting Efficiency of RNAi
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1.
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The shRNA vector elements: like loop structure, thermodynamic properties of the shRNA hairpin;
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2.
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Target gene: like secondary structure, the surrounding sequences;
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Experimental system: such as, target cells: for hard-to-transfect cells(primary cells), our virus products or virus packaging services will help you solve the problem of poor transfection efficiency; the cell state and confluence: for transfection to be successful, the cells must be in a good cell state; the observation time, depends on the half-life of the target protein; the transfection reagents and transfection steps will also affect the transfection efficiency;
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4.
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The off-target effects;
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5.
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The compensation responses: mask RNAi-specific effects.
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