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Small interfering RNA targeting penaeus monodon TRIM9 gene and application of small interfering RNA

A technology of penaeus monodon and small interference, applied in the field of small interference RNA, can solve problems such as outbreaks, and achieve the effects of ensuring economic benefits, inhibiting replication, and improving survival rate

Active Publication Date: 2021-04-30
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, studies have found that white spot virus (WSSV) can hijack the TRIM9 gene of Litopenaeus vannamei for its own use, inhibiting the NF-κB pathway and the production of downstream antimicrobial peptides, leading to large-scale shrimp disease outbreaks

Method used

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  • Small interfering RNA targeting penaeus monodon TRIM9 gene and application of small interfering RNA
  • Small interfering RNA targeting penaeus monodon TRIM9 gene and application of small interfering RNA
  • Small interfering RNA targeting penaeus monodon TRIM9 gene and application of small interfering RNA

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Embodiment 1

[0027] Cloning of embodiment 1 Penaeus monodon TRIM9 gene and its expression analysis under Vibrio parahaemolyticus infection

[0028] 1. Total RNA Extraction and cDNA Synthesis

[0029] According to the requirements of the TRIZOL reagent (Invitrogen, shanghai) instructions, extract healthy Penaeus monodon (about 25g) muscle (muscle), heart (heart), hepatopancreas (hepatopancreas), brain (brain), stomach (stomach), gill (gill), Total RNA of blood cells (hemocytes) and intestinal tract (intestine); according to the instructions of the PrimeScript reverse transcriptase kit (TaKaRa, Dalian), the above total RNA was reverse-transcribed to synthesize cDNA, and stored at -80°C for future use.

[0030] 2. Amplification and bioinformatic analysis of TRIM9 gene open reading frame (open reading frame, ORF) in Penaeus monodon

[0031] Using primers PmTRIM9-F (5'-ATGGAGGAGGAGCTGCGG-3') and PmTRIM9-R (5'-CTATGTTTTAGCAGCAATAGGAGTC-3'), the open reading frame sequence of P. monodon TRIM9 was ...

Embodiment 2

[0036] Example 2 Preparation of small interfering RNA sequence (TRIM9-siRNA) targeting the TRIM9 gene of Penaeus monodon and its application in the process of resisting Vibrio parahaemolyticus

[0037] 1. Preparation of the small interfering RNA sequence (TRIM9-siRNA) working solution targeting the TRIM9 gene of Penaeus monodon:

[0038] 1.1. According to the TRIM9 open reading frame sequence (GenBank accession No.MW18461) cloned in the present invention, use BLOCK-iT TM RNAi Designer online program (https: / / rnaidesigner.thermofisher.com / rnaiexpress / sort.do) designed siRNA sequences, including TRIM9-siRNA sense strand sequence as shown in SEQ ID NO.1, antisense strand sequence as SEQ ID NO. 2.

[0039] Sense strand sequence: 5'-GCUCCGUUCUGACGCCGACAGACG-3'

[0040] Antisense strand sequence: 5'-CGUCUGUCGGCGUCAGAACGGAGC-3'.

[0041] After the sequence was designed, entrust Shanghai Gemma Company to synthesize and synthesize separately. The TRIM9-siRNA lyophilized powder synt...

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Abstract

The invention discloses a small interfering RNA targeting a penaeus monodon TRIM9 gene, wherein the small interfering RNA has a sense strand sequence of 5'-GCUCCGUUCUGACGCCGACAGACG-3' and an antisense strand sequence of 5'-CGUCUGUCGGCGUCAGAACGGAGC-3'. The invention also discloses an application of the small interfering RNA. The TRIM9-siRNA disclosed by the invention can obviously inhibit the expression of the penaeus monodon TRIM9 gene, promote the expression of various antibacterial peptide genes, inhibit the replication of vibrio parahaemolyticus in penaeus monodon, and improve the survival rate of the penaeus monodon under the infection of the vibrio parahaemolyticus.

Description

technical field [0001] The present invention relates to a small interfering RNA (small interfering RNA, siRNA), in particular to a small interfering RNA targeting the TRIM9 gene of Penaeus monodon. At the same time, the present invention also relates to its use in resisting Vibrio parahaemolyticus infection. Background technique [0002] Small interfering RNA (small interfering RNA, siRNA) is a double-stranded RNA with a length of 20-25 nucleotides, which has many different uses in biology. At present, it is known that siRNA is mainly involved in the phenomenon of RNA interference (RNAi) in organisms, and regulates the expression of functional genes in a specific silencing manner. In recent years, RNA interference technology has made great progress in the treatment of bacteria, viruses and other diseases, especially in the treatment of human AIDS, hepatitis B and other diseases. The latest research shows that siRNA targeting HIV structural protein gene and long terminal re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/713A61P31/04
CPCA61K31/713A61P31/04C12N15/113C12N2310/14C12N2320/30
Inventor 赵超邱丽华彭超范嗣刚王鹏飞闫路路
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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