Cell-based shRNA/siRNA Validation Service

Creative Biogene has established fully-matured RNAi system, ranges from design, siRNA synthesis/shRNA construction, transfection, expression evaluation to functional testing. On this basis, we offer cell-based RNAi service to verify the silencing effects of siRNA transient transfection and/or shRNA incorporation and stable expression. Creative Biogene has the ability to accurately evaluate the silencing activities of shRNA and siRNA, and assist you to screen shRNA/siRNA with high knockdown efficiency.

RNA interference (RNAi) is the process by which the expression of a target gene is effectively silenced or knocked down by the selective inactivation of its corresponding mRNA by double-stranded RNA (dsRNA). The silencing mechanisms can either lead to the degradation of a target mRNA, as induced by small interfering RNAs (siRNAs) or short hairpin RNAs (shRNAs), or the suppression of translation of specific mRNAs, as induced by microRNA (miRNA). RNAi technology allows researchers to analyze gene function, determine disease pathways and identify potential drug targets.

Figure 1. Mechanism of RNA interference.

There are many algorithms that predict most effective siRNA and shRNA sequences, however experimental in vitro test of such sequences usually demonstrate that less than 30% of the constructs tested provide high level of gene silencing (over 80% target mRNA reduction). Thus, experimental evaluation of the activities of shRNA/siRNA still play a key role in expanding its application in biomedical and gene therapy fields.

Workflow of Cell-based Knockdown Validation

• siRNA/shRNA design and prediction via various algorithms
• siRNA oligo synthesis and modification (optional)
• cloning 4-6 predicted shRNA in suitable vector (optional)
• siRNA transient transfection, or shRNA incorporation and stable expression
• Quantification of target gene silencing via RT-qPCR and western blot

Professional team at Creative Biogene compare the efficiency of shRNA/siRNA candidates in a cell model of interest to you; analyzing various factors to select the best one for knockdown of your target gene. We guarantee the knockdown of >70% at mRNA level in the tested cells by screening 4 shRNA clones. While testing silencing efficacy, we will take all details into consideration, such as overall transfection efficiency of certain cell lines and use of associated positive/negative controls. For more information, please feel free to contact us.

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