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SLE-related circular RNA hsa_circ_0025843 and application thereof

A rna-circ-fgd4, circular technology, applied in the fields of molecular biology and immunity, can solve the problems of unknown key mechanisms, and achieve the effect of promoting proliferation and secretion of IL-2

Active Publication Date: 2021-05-25
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a large number of studies have shown that genetics, epigenetics, environmental factors and immunological mechanisms are involved in the occurrence and development of systemic lupus erythematosus, the key mechanism is still unknown.

Method used

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  • SLE-related circular RNA hsa_circ_0025843 and application thereof
  • SLE-related circular RNA hsa_circ_0025843 and application thereof
  • SLE-related circular RNA hsa_circ_0025843 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036]Example 1: Discovery and detection of CIRC_FGD4 in clinical samples

[0037]SeeFigure 1 to 3 ,

[0038](1) High-throughput sequencing technology screens the cycloid RNA CIRC-FGD4 in peripheral blood of SLE patients

[0039]1. Collection of experimental groups and blood samples

[0040]A total of 30 patients with SLE (aged 39.5 ± 12.1 years), the First Affiliated Hospital of Wenzhou Medical University, the rheumatoid immunology of the Second Affiliated Hospital of Wenzhou Medical University, 2016.17.5 months, 2017. May. The selected cases comply with the SLE diagnostic criteria revised by the US Rheumatology Association (ARA) in 1997. In patients with glucocorticoids and immunosuppresses, blood specimens collection and systemic lupusdisease activity index (SYSTEX, SLYTHEMATISDISEASE Activity Index, Sledai) statistics were completed. The normal control group was 30 healthy volunteers (aged 35.8 ± 10.0 years old), no life-free disease or immunosuppressive treatment history. The control group...

Embodiment 2

[0069]Example 2: Stability Experiment of CIRC_FGD4

[0070]SeeFigure 4 to 5 ,

[0071]1. Study the stability of CIRC_FGD4 on the oligogenic enzyme (RNASE R)

[0072](1) RNASE R experiment

[0073]The CD3 + T cells with SLE patients and health volunteers were taken as the study, and RNA was extracted by RNAase R experiment, divided into RNAase R +, RNAase R-group.

[0074]Reaction system: Storage Buffer: 50mm Tris-HCl (pH 7.5), 100mm NaCl, 0.1 mm EDTA, 1MM DTT, 0.1%X-100, 50% Glycerol 10X Reaction Buffer: 200mm Tris-HCl (pH 8.0), 1M KCL, 1MM MGCL2

[0075]Configure the RNAase R + Reaction System (Ice on Ice), see Table 2.

[0076] Reagent name Usage amount RNA > 5UG 10x Reaction Buffer 5ul RNAase R 3-4 ul / ng RNase-Free H2O TO 50UL

[0077]Table 2

[0078]The above reaction liquid was sufficiently mixed, placed in a 37 ° water bath for 15 min.

[0079](2) Reverse transcription reaction after RNAase R experiment

[0080]The first chain cDNA synthesis reaction liquid is quantified in 1 ug.

[0081]The following mi...

Embodiment 3

[0099]Example 3: Positioning of CIRC_FGD4

[0100]SeeFigure 6 ,

[0101]The Fish probe sequence of CIRC_FGD4 is as follows, see Table 5.

[0102]

[0103]table 5

[0104]1) Probe dilution: NMOLE / OD260 = 4.17, 41.7 μl of sterilized DEPC water was added to each OD probe dry powder, and the concentration of about 100 μm was obtained.

[0105]2) Formulate the probe mixture: 70 μl of Buffer E, 2μL probe, 28 μl of DEPC water, end volume is 100 ul

[0106]3) Smertrane: The slide of about 20 ul of T cell suspension is dropped on the -20 degree slide, and also take a slide as a push plate, when the cell drop is evenly attached between two pieces, The angle of the two slides is 30 to 40 degrees, and the pusher is slowly uniformly pushed into the right side of the slide from the left side of the cell drop, and the uniform cell membrane is introduced, and the slide is placed on the alcohol lamp. Grilled into a white film.

[0107]4) Place the grilled cell smear in 10 cm Dish and dripped 100 μl of 0.1% buffer a for 1...

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Abstract

The invention discloses a circular RNA circ-FGD4 and an application thereof. The nucleotide sequence of the circular RNA circ-FGD4 gene is shown as SEQ ID NO: 1; in addition, the invention provides a pharmaceutical composition, a systemic lupus erythematosus diagnostic kit and application of the circ-FGD4 gene as a target gene in preparation of drugs for treating systemic lupus erythematosus. The circ-FGD4 gene and the expression product thereof are used as markers for diagnosing systemic lupus erythematosus, so that the systemic lupus erythematosus can be diagnosed more accurately and quickly, and the circ-FGD4 gene and the expression product thereof are used as target genes for preparing medicines for treating systemic lupus erythematosus to provide a new therapeutic target and a new therapeutic approach for treating systemic lupus erythematosus.

Description

Technical field[0001]The present invention relates to the field of molecular biology and immunization, and more particularly to a SLE-related cyclic RNA HSA_CIRC_0025843 and its applications thereof.Background technique[0002]SystemicLuPusertHematosus, SLE) has been considered a typical hypersensitive state in many years. This state is mediated by an inflammatory process after organ specific deposition of immune complexes. Along with complementary system The activation of the inflammatory pathway. Among the SLE patients, there is an immunocyte dysfunction, particularly abnormal activated T / B lymphocytes, resulting in a large number of autoantibodies, and attacks their own tissue organs cause damage. During systemic lupus erythematation, T and B lymphocytes play a crucial role, especially the recirculation of T lymphocytes, and enhance immune responses, further playing cellular immunity and immune regulation. Function, strengthen autoimmunity and promote the immune exclusion of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113C12N15/11A61K48/00A61K31/7088A61P29/00A61P37/02
CPCC12Q1/6883A61K31/7088A61P29/00A61P37/02C12Q2600/178C12Q2600/158
Inventor 叶辉万乐予向敏王雪汪子晗褚晓颖胡煊煊李美琪杜邦张玉桑侃如
Owner WENZHOU MEDICAL UNIV
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