circRNA Pull-down Assay

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circRNA Pull-down Assay

IntegrateRNA is an outstanding biotechnology company with years of experience in RNA-protein interaction analysis. We offer customized circRNA pull-down assay service to help you validate the interaction of circRNA-miRNA and circRNA-RBP. We always work together with you to solve specific challenges and chase highest-level customer satisfaction.

Circular RNA (circRNA) is recently discovered as a special novel type of endogenous noncoding RNA (ncRNA), and attracts considerable attention due to its powerful functions and unique properties. It’s reported that some circRNAs contain several miRNA binding sites and act as a sponge to impact the bio-function of miRNA. In addition, circRNA can serve as a protein decoy which harbors binding sites for a single or multiple proteins and regulate gene expression. Thus, the interaction analysis of circRNA-miRNA and circRNA-RBP is crucial to reveal the biological function and mechanism of circRNA.

RNA pull-down is an in vitro technique for detection of protein-RNA interactions using labeled RNA as bait. Currently, biotinylated probes based on back-splicing junction sequence are widely applied to circRNA pull-down assay. However, due to the probe sequence overlap with the often more abundant linear host RNA, it is difficult to completely exclude the interference of the host linear RNA. IntegrateRNA introduces the up-to-date circRNA pull-down technique to help you identify the interaction between miRNA/RBP and circRNA.

Strategy of circRNA pull-downFigure 1. Strategy of circRNA pull-down

As shown, circRNA pull-down assay depends on the principle that capture protein has a strong affinity to tagged RNA, and can effectively and specifically pull down circRNA–miRNA/protein complexes. The eluted proteins are then detected by western blotting or mass spectrometry while the pull-down RNAs are detected by northern blot or through RT-PCR analysis.

Workflow of circRNA Pull-down Assay

  • Construction of tagged circRNA expression vector
  • Construction of captured protein expression vector
  • Co-transfection
  • Inducing the trapped protein expression
  • Pull-down assay using labeled antibody
  • Elution
  • Detection of eluted proteins and RNAs

Advantages

  • Avoiding interference of the host linear RNA
  • Capturing target molecular effectively and specifically
  • Suitable for low abundance expression circRNAs

Applications

  • Identifying miRNAs and proteins that interacted with circRNA
  • High-throughput screening of potential interaction molecules

With many years of experience in RNA research, IntegrateRNA introduced the most up-to-date circRNA pull-down technique to help you identify the interaction between miRNA/RBP and circRNA. Each project is fully customized to meet your scientific needs, and our experienced team can provide flexible solutions for any individual request. We guarantee our clients the most reliable and efficient research services with faster turnaround time and lower prices. If you have any questions, please feel free to contact us.

For research use only. Not intended for any clinical use.
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