CircRNA overall project

TOPBIOTECH can provide "high-throughput sequencing + bioinformatics analysis + downstream functional verification" one-stop service for circRNA research.

Introduction

Circular RNA (or circRNA) is a type of RNA which, unlike the better known linear RNA, forms a covalently closed continuous loop, i.e., in circular RNA the 3' and 5' ends normally present in an RNA molecule have been joined together. Because circular RNAs do not have 5' or 3' ends, they are resistant to exonuclease-mediated degradation and are presumably more stable than most linear RNAs in cells.

Certain circular RNAs have many miRNA binding sites, some circular RNAs acting as miRNA sponges, block the inhibitory effect of miRNA on its target genes. Through base complementary pairing, circRNA directly regulates other RNA levels, circRNA can also bind to proteins to regulate protein activity.

circRNA research plan

Technical process

      1、  Screening circRNA (looking for differentially expressed circRNA): RNA-seq, microarray, bioinformatics analysis;

2、  circRNA identification and localization: RT-PCR, Sanger sequencing, Northern blot analysis, RNA-Fish;

3、  circRNA expression analysis: cell level, tissue level and clinical sample verification;

4、  In vitro function assessment of circRNA: overexpression or interference of circRNA, cell transfection, cell scratch test, transwell migration and invasion test, etc;

5、  In vivo function assessment of circRNA: overexpression or interference of circRNA,  construction of xenotransplantation model, immunohistochemistry, RT-QPCR, WB, etc;

6、Mechanism research: sequence prediction and alignment, design of microRNA probe and circRNA probe, RNA pulldown, RT-QPCR and dual-Luciferase reporter assay system to determine the binding site.