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Analysis/diagnosis method utilizing RNA modification

A technology for information analysis and status, applied in the field of analysis and diagnosis using RNA modification, which can solve problems such as insufficient prediction of early cancer

Pending Publication Date: 2020-11-27
OSAKA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to analyze various biological states including diseases, it is attempted to correlate information such as DNA variation, mRNA expression level, protein expression level, etc. more

Method used

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  • Analysis/diagnosis method utilizing RNA modification
  • Analysis/diagnosis method utilizing RNA modification
  • Analysis/diagnosis method utilizing RNA modification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0630] (Example 1) Methylation analysis of microRNA

[0631] In this example, methylation analysis of microRNA was carried out. The details are as follows.

[0632] MicroRNA 200-c-5p (human sequence, SEQ ID NO: 11) synthesized to contain methylated adenine and methylated cytosine was dissolved in RNase Free ultrapure water, the concentration was confirmed by absorbance measurement, and adjusted to 1 pmol / μL. Using 1 μL of the microRNA aqueous solution, mass spectrometry was performed with a MALDI-type mass spectrometer according to the above-mentioned protocol. To confirm the internal sequence, the RNA was degraded by ammonia treatment (5' → 3'). In addition, measurement using in source decay (ISD) was carried out for the observed precursor ion ( figure 1).

[0633] Mix synthetic oligo DNA (complementary strand of human 369-3p, SEQ ID NO: 12) with a sequence complementary to microRNA 369-3p and its antisense synthetic DNA (SEQ ID NO: 13) in equimolar amounts, heat and t...

Embodiment 2

[0637] (Example 2) Analysis of cancer using RNA modification

[0638] In this example, the influence of cancer on RNA modification was analyzed to verify that cancer can be detected or diagnosed by using RNA modification.

Embodiment 2-1

[0639] (Example 2-1: RNA modification analysis of cell lines)

[0640] Human pancreatic cancer cell lines BxPC3, Panc10.5, PSN1 and Capan2 were obtained from American Type Culture Collection (Manassas, VA, USA). For these four lines, methylated miRNAs were analyzed by RIP-Seq using an anti-m6A antibody, and 63 types of methylated miRNAs common to the four pancreatic cancer cell lines were found, as shown in the table below.

[0641] [Table 10]

[0642]

[0643] Methylation of these miRNAs may be useful in the determination of cancer (eg, pancreatic cancer).

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Abstract

The present application provides a novel method for analyzing a biological condition or a medical condition. It is now found that information about the modification of RNA can be used for the analysisof a biological condition or a medical condition. On the basis of this finding, the present invention provides a novel method for analyzing a biological condition or a medical condition. According tothe present invention, it becomes possible to analyze various biological conditions or medical conditions including diseases, and it also becomes possible to satisfactorily predict cancer (e.g., pancreatic cancer, colorectal cancer, stomach cancer) in an early stage.

Description

technical field [0001] The invention relates to a feature analysis method and classification of biological objects. More specifically, the present invention relates to methods for classifying and characterizing biological objects based on modification information on RNA. Background technique [0002] Like deoxyribonucleic acid (DNA), ribonucleic acid (RNA) is a molecule that carries information possessed by living things. It is well known that DNA undergoes modification such as methylation to control its function, and in recent years it has been reported that RNA is also modified. [0003] In order to analyze various biological states including diseases, it is attempted to correlate information such as DNA variation, mRNA expression level, protein expression level, etc. more. [0004] prior art literature [0005] non-patent literature [0006] Non-Patent Document 1: Pagliarini DJ, Cell Metab.2016Jul 12; 24(1):13-4.doi:10.1016 / j.cmet.2016.06.018. Contents of the inven...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12M1/00C12Q1/6806C12Q1/6872C12Q1/6888G01N33/50C12N15/09C12N15/10
CPCC12Q1/6806C12Q1/6869C12Q1/6872C12Q1/6886C12Q2600/178C12N15/1013C12Q1/6827C12Q2600/106C12Q2600/154
Inventor 石井秀始今野雅允森正树
Owner OSAKA UNIV
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