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Exosome secreted by lncRNA gene modified cell and application thereof

A technology of exosomes and cell lines, applied in the field of cell biology, can solve problems such as deficiencies

Active Publication Date: 2022-02-18
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the effects of long non-coding RNA-regulated exosomes on macrophage cytokines.

Method used

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  • Exosome secreted by lncRNA gene modified cell and application thereof
  • Exosome secreted by lncRNA gene modified cell and application thereof
  • Exosome secreted by lncRNA gene modified cell and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Construction of lncENAF-overexpressing lentiviral vector

[0037] The research group has kept the pMD-T18-lncENAF plasmid and strain. In the early stage, lncENAF (the nucleotide sequence of lncENAF is shown in SEQ ID NO: 1) was designed by designing homology arm primers (restriction sites: XbaI and SalI). Linked to the pCDH-GFP plasmid to form the pCDH-GFP-lncENAF plasmid (see the plasmid map image 3 , the nucleotide sequence of pCDH-GFP-lncENAF is shown in SEQID NO: 2), and the homology arm primers are shown in the accompanying drawings (see Table 1), and the specific construction steps are as follows:

[0038] Table 1 Homology Arm Primer Sequence

[0039]

[0040] (1) PCR amplification of lncEANF: use PCR to amplify lncEANF, the amplification template is pMD-T18-lncENAF, and the amplification system is as follows:

[0041] Table 2 Amplification system

[0042]

[0043] (2) Weigh 0.5g of agarose powder into a clean Erlenmeyer flask, add 50mL of TAE buffer, h...

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Abstract

The invention discloses an exosome secreted by an lncRNA gene modified cell and application, and belongs to the field of cell biology. The exosome is secreted by an HEK-293T cell strain obtained through genetic engineering, the HEK-293T cell strain stably expresses lncENAF, and the nucleotide sequence of the lncENAF is as shown in SEQ ID NO: 1. The exosome can effectively inhibit generation of lipopolysaccharide-induced macrophage cytokines, is a potential cytokine inhibitor, and provides a new direction for cytokine storm and treatment of autoimmune diseases.

Description

technical field [0001] The present invention relates to the field of cell biology, in particular to an exosome secreted by lncRNA gene-modified cells and its application. Background technique [0002] Long non-coding RNA (lncRNA) is a kind of non-coding sequence with a transcription length of more than 200nt, which rarely or cannot encode protein because of the lack of an effective open reading frame. Due to the shallow research on lncRNA before, it is believed that it does not have any biological function, but exists only as a by-product of the transcription process. With the continuous development of molecular biology sequencing technology, it is found that it can regulate genes from different levels such as epigenetic regulation, transcriptional regulation and post-transcriptional regulation. Therefore, the regulatory role of long non-coding RNA is being paid more and more attention and researched. [0003] Exosomes refer to extracellular vesicles with a particle size o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/867C12N5/10A61K35/17A61P7/00A61P37/02
CPCC12N15/1136C12N15/86C12N5/0686C12N5/0625A61K35/17A61P7/00A61P37/02C12N2740/15043C12N2510/02C12N2502/1157Y02A50/30A61K35/22C12N5/0645A61K9/5068C12N2740/16043C12N2310/111C12N2330/50C12N2502/256C12N15/113
Inventor 李伟刘林胡旭东
Owner WENZHOU MEDICAL UNIV
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