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

A technology of expression abundance and gene chip, applied in the field of detection of circular RNA expression abundance, can solve the problem of low research throughput and achieve the effect of high-throughput detection

Inactive Publication Date: 2016-07-27
上海伯豪生物技术有限公司
View PDF1 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Probe, gene chip and method for detecting expression abundance of circular RNA
  • Probe, gene chip and method for detecting expression abundance of circular RNA
  • Probe, gene chip and method for detecting expression abundance of circular RNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0028] In order to have a more specific understanding of the technical content, characteristics and efficacy of the present invention, the specific method for detecting the expression abundance of circular RNA of the present invention is described in detail as follows in conjunction with the accompanying drawings:

[0029] 1. Design chip probe sequence

[0030] According to the special splicing pattern of circular RNA-there is a specific reverse splicing site where the 5'end of the donor exon and the 3'end of the acceptor exon are connected end to end (backsplicing, linear RNA does not have this site) , The circular RNA probe is designed on the 60nt chip probe sequence located at the reverse splicing site, which can specifically detect circular RNA in the sample.

[0031] The circular RNA detected in this example is hsa_circ_0034688, which is formed by reverse splicing from the fourth exon to the fifth exon of a gene named OIP5. According to the 5'end sequence of the circular RNA:

...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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 ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6837C12Q1/6876C12Q2600/158C12Q2600/178C12Q2525/307
Inventor 黄建锋张晓娜邓俊豪蒋敏肖华胜
Owner 上海伯豪生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap