Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Circular RNA and application thereof

A circular, nucleotide sequence technology, applied in the field of circular RNA

Active Publication Date: 2020-07-07
THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although osimertinib is more effective than first- and second-generation EGFR-TKIs such as gefitinib and afatinib to a certain extent, drug resistance is still inevitable

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
  • Circular RNA and application thereof
  • Circular RNA and application thereof
  • Circular RNA and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Detect the expression of circ_0058040 in tissues and cells

[0030] (1): The expression of circ_0058040 in normal lung tissue and lung cancer tissue:

[0031] 1. Chip preparation and analysis: Prepare normal lung tissue and lung cancer tissue samples according to the requirements of Shanghai Bohao Company, and submit them to Bohao Company for chip preparation and analysis. The chip contains the latest versions of many circRNAs databases and tens of thousands of circRNAs detection probes Needle. The detection principle of the chip is based on the length and structural characteristics of the circRNA sequence, the reverse transcription method of random primers is used to introduce the T7 promoter, and then the RNA is amplified by the in vitro transcription-mediated linear amplification method. The RNA linear amplification method uses Sample labeling in microarray chip analysis. The DNA obtained by reverse transcription of the linear amplification product cDNA is then flu...

Embodiment 2

[0047] Construct circ_0058040 lentiviral interference vector to transfect cells:

[0048] Design a specific interference sequence for circ_0058040, its sequence is (5'to 3'): AGATGAAAGGCAATACAAAGGAACGGAAdTdT, see Seq NO.2, take the sequence fragment of the interference GAPDH gene as a control, and insert it into the lentiviral vector, see Figure 8 . Infect the non-small cell lung cancer cell line A549 with the packaged lentiviral vector to down-regulate the expression of circ_0058040. After 48 hours, use G418 to screen the stably transfected cell line and name it A549 / sh-circ_0058040 (the control is A549 / NC) ,

Embodiment 3

[0049] Example 3: Cell Proliferation Experiment and Colony Formation Experiment

[0050] 1. Colony formation experiment:

[0051] aExperimental method:

[0052] 1) Cell inoculation: digest monolayer cultured cells with 0.25% trypsin, prepare a single A549 / sh-circ_0058040 (control is A549 / NC) cell suspension with 1640 culture medium containing 10% fetal bovine serum, and use 500 cells were seeded in a 6-well culture plate;

[0053] 2) Put it in a cell incubator, change the medium every 4 days, and cultivate for 2 weeks;

[0054] 3) When clones visible to the naked eye appear in the culture dish, the culture is terminated. Discard the supernatant and gently wash twice with PBS;

[0055] 4) Add 1ml of pure methanol or 1:3 acetic acid / methanol, fix for 15 minutes;

[0056] 5) Discard the methanol fixative, add 1ml of 0.1% crystal violet staining solution to stain for 15 minutes, then slowly wash away the staining solution with PBS, and air dry;

[0057] b. Results determinat...

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 belongs to the field of genetic engineering and particularly relates to an application of a circular RNA circ-0058040 to preparation of a drug for treating a non-small cell lung cancer.Proliferation, apoptosis and drug susceptibility of a non-small cell lung cancer cell are influenced by changing expression of the circular RNA circ-0058040; the application illustrates that the proliferation of the non-small cell lung cancer cell can be inhibited by the expression of the circular RNA circ-0058040; and the susceptibility of the non-small cell lung cancer cell to a targeted drug can be enhanced.

Description

technical field [0001] The invention belongs to circular RNA, in particular to the application of circular RNA circ_0058040 in the preparation of drugs for treating non-small cell lung cancer. Background technique [0002] Circular RNA (circular RNA, circ RNA) is a new type of RNA that is different from traditional linear RNA and characterized by covalently closed loops, and is more stable and conserved than linear RNA. It was first discovered in plant viruses by Nobel laureate Sanger in 1976. Limited by experimental techniques and research methods, circRNA was considered to be a low-abundance RNA molecule formed by mis-splicing of exons in the 30 years after its discovery. In recent years, benefiting from the rapid development of high-throughput sequencing technology and bioinformatics, many circRNAs have been discovered in different species. [0003] circRNA has become a new hotspot in tumor research because of its potential role in carcinogenesis and tumor suppression p...

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
IPC IPC(8): C12N15/113A61K31/7088A61P35/00A61P11/00
CPCC12N15/113A61P35/00A61P11/00C12N2310/532C12N2310/113C12N2310/14
Inventor 王朝霞陈臻瑶陈昕顾婧瑶
Owner THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com