Probe, gene chip and method for detecting expression abundance of circular RNA

Abstract

The invention discloses a probe for detecting expression abundance of circular RNA. The probe includes backsplicing sites at the 5' terminal of a donor exon and at the 3' terminal of an accepter exon of the circular RNA. The invention also discloses a gene chip containing the probe, and a method for detecting the expression abundance of the circular RNA by virtue of the gene chip, wherein the method mainly comprises the following steps: 1) extracting and purifying total RNA of a to-be-detected sample; 2) removing ribosome RNA; 3) enriching circular RNA; 4) conducting linear amplification to the enriched circular RNA and labeling fluorescence; 5) hybridizing a fluorescence-labeled product with a chip probe; and 6) washing and scanning the chip, and analyzing data. By designing the probe and the gene chip which are capable of achieving specific detection of the expression abundance of the circular RNA, the rapid and high-throughput detection of the expression abundance of the circular RNA can be achieved, so that shortcomings in an existing circular RNA detection technology are overcome.

Description

Technical field [0001] The present invention relates to the field of biology, in particular to the detection of the expression abundance of circular RNA. Background technique [0002] CircularRNA (circRNA) is a new type of RNA molecule that is characterized by a covalent closed loop and widely exists in eukaryotes. circRNA is derived from the exon or intron regions of genes, and is abundant in mammalian cells. The formation of circRNAs is different from the standard splicing mode of linear RNA. The 5'end of the donor exon and the 3'end of the acceptor exon are connected end to end to form a backsplicing site (backsplicing). . The existing circRNA formation models mainly include the following (see figure 1 Shown): [0003] (1) “lariat-drivencircularization” or “exonskipping”, such as figure 1 Shown in A; [0004] (2) "intron-pairing-drivencircularization" or "directbacksplicing", such as figure 1 Shown in B; [0005] (3) The formation pattern of circular intron RNAs (ciRNAs), such ...

AI Technical Summary

Problems solved by technology

At present, the commonly used technical method for detecting the expression abundance of circular RNA is real-time PCR, but this method has the disadvantage of low research throughput, and the research on circRNA has just started, and a high-throughput reliable detection technology is urgently needed to meet the research needs.

Images