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circptpn22 and its application as a marker of systemic lupus erythematosus

A lupus erythematosus, systemic technology, applied in the biological field, can solve problems such as imperfections

Active Publication Date: 2021-07-06
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing circRNA database—circBASE is not perfect, and there are still many unrecorded circRNAs. Therefore, we used the third-generation sequencing technology—RNA-seq to sequence the peripheral blood mononuclear cells (PBMCs) of SLE patients and normal people. Further search for specific circRNAs that may act on diseases

Method used

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  • circptpn22 and its application as a marker of systemic lupus erythematosus
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  • circptpn22 and its application as a marker of systemic lupus erythematosus

Examples

Experimental program
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Embodiment 1

[0015] Example 1 proves the existence of circPTPN22

[0016] A. In the high-throughput sequencing data, sort the log2FC value from high to low to find a relatively stable circRNA (log2FC≥1.0and FDR≤0.05), through comparison, select a circRNA whose parent gene is PTPN22, and obtain its sequencing result The sequence information and its cyclization site are named circPTPN22, and the sequence of circPTPN22 is shown in SEQ ID NO.1.

[0017] B. Qrt-PCR

[0018] details as follows:

[0019] 1. The difference between circRNA and ordinary RNA lies in the circular cleavage site (joint), so to specifically prove the existence of a new circRNA, it is necessary to design primers across the circular cleavage site. By the end-to-end splicing method, primers across the cleavage site were designed in Primer5 software: 5'-GGAGTCCACTGGCGTCTTC-3' (upstream primer) and GCTGATGATCTTGAGGCTGTTG-3' (downstream primer), such as SEQ ID NO.2 and SEQ ID NO.3 As shown, use Oligo7 to verify the primers....

Embodiment 2

[0024] Example 2 circPTPN22 can be one of the markers of SLE

[0025] A. Collect 5ml of peripheral blood from SLE patients and healthy subjects, separate PBMCs, and extract RNA;

[0026] B. qrt-pcr large-scale verification of the expression of circPTPN22 in patients and normal people;

[0027] C. Using the analysis software CFX manager and SPSS 22.0 to analyze the correlation between circPTPN22 and SLE.

[0028] details as follows:

[0029] 1. Large-scale collection of peripheral blood (5ml per person) from healthy people and SLE patients, collected in EDTA vacuum anticoagulation tubes (BD Company, USA).

[0030] 2. Absorb peripheral blood, slowly add it to the centrifuge tube that has been added with lymphocyte separation solution (3ml, Tianjin Haoyang), keep the liquid surface boundary line clear, centrifuge at 2000r / min for 20 minutes, take it out slowly, and absorb the middle buffy coat layer to another Into a new centrifuge tube, add 5ml of PBS (biosharp), centrifuge a...

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Abstract

The present invention relates to circPTPN22 and its application as a marker of systemic lupus erythematosus. The present invention first confirmed the existence of a new type of circRNA, namely circPTPN22, and confirmed its correlation with SLE in a large-scale patient population study. The first confirmed circRNA marker associated with SLE laid the foundation for the in-depth study of circRNA in SLE.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to circPTPN22 and its application as a systemic lupus erythematosus marker. Background technique [0002] circRNA is a circular closed RNA that exists widely and diversely in various biological cells. In recent years, with the maturity of high-throughput sequencing technology, more and more circRNAs have been discovered, and its characteristics—stable, specific, broad, conserved, and functional diversification—suggest that it can be used as an important disease markers. Systemic lupus erythematosus (SLE) is a typical autoimmune disease that can lead to the involvement of various organs and systems throughout the body, such as the skin, kidneys, lungs, etc. The clinical manifestations are diverse and individual differences are large, and it is still incurable , Most patients rely on glucocorticoids for maintenance treatment all year round. Clinically, SLE is diagnosed mainly by clinical...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12N15/11C12N2310/532C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 倪兵苗青青仲志婷
Owner ARMY MEDICAL UNIV