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siRNA of schistosoma japonicum SjELAV-like 1 genes and application thereof

A technology for schistosomiasis and schistosomiasis, which is applied in the fields of molecular biology and biomedicine, and can solve problems such as lack of in-depth research.

Active Publication Date: 2018-11-13
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no in-depth study on the ELAV-like 1 gene in Schistosoma japonicum

Method used

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  • siRNA of schistosoma japonicum SjELAV-like 1 genes and application thereof
  • siRNA of schistosoma japonicum SjELAV-like 1 genes and application thereof
  • siRNA of schistosoma japonicum SjELAV-like 1 genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 siRNA screening

[0021] 1. Method steps

[0022] 1.1 Preparation of siRNA molecules and primers for real-time quantitative PCR of SjELAV-like 1 gene

[0023] According to the SjELAV-like 1 (MG515727) gene sequence submitted to NCBI (http: / / www.ncbi.nlm.nih.gov / ), two pairs of dsRNA were designed and synthesized through analysis and comparison. Simultaneously, a pair of irrelevant control dsRNA (NCdsRNA) was synthesized as:

[0024] sense: 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID NO.5),

[0025] Anti-sense: 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID NO.6);

[0026] The dsRNA small molecules of two pairs of Schistosoma japonicum ELAV-like 1 genes are:

[0027] S1dsRNA: sense: 5'-GCUAUACCUGUACGUAUUATT-3' (SEQ ID NO.1),

[0028] Anti-sense: 5'- UAAUACGUACAGGUAUAGCTT -3' (SEQ ID NO.2), the target sequence of the first pair of siRNAs is 648-664bp (UAUACCUGUACGUAUUA);

[0029] S2dsRNA: sense: 5'-GCGACAGUGAAUUUACCUUTT-3' (SEQ ID NO.3),

[0030] Anti-sense: 5'- AAGGUAAAUUC...

Embodiment 2

[0037] Example 2 Interference in mice

[0038] 1. Method steps

[0039] 1.1 Long-term interference test of siRNA in vivo

[0040] 1.1.1 siRNA long-term interference

[0041] Eight BALB / c mice were divided into 2 groups, 4 mice in each group, and the mice were infected according to the above method. On day 14 after infection, the siRNA with the best interference effect was selected, and the mice were interfered with in the same way, every 4 d interfered once, a total of 7 times, and the worms were collected on the 41st day.

[0042] 1.1.2 Real-time quantitative PCR verification of RNA interference effect in vivo

[0043] The total RNA of each group of worms was extracted with Trizol kit, the genome was digested, and reverse transcribed into cDNA, which was used as a template for real-time quantitative PCR analysis.

[0044] 1.1.3 Liver egg count

[0045] Collect the liver and weigh it. Add 20mL of PBS to the liver tissue, mix it with a homogenizer, add an equal volume of 10%...

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Abstract

The invention discloses siRNA of schistosoma japonicum SjELAV-like 1 genes. The siRNA has nucleotide sequences shown as SEQ ID NO.3 and SEQ ID NO.4. The invention further discloses the application ofthe siRNA of schistosoma japonicum SjELAV-like 1 genes. The siRNA of schistosoma japonicum SjELAV-like 1 genes disclosed by the invention is capable of obviously inhibiting transcription of the schistosoma japonicum SjELAV-like 1 genes and obviously reducing the number of liver loaded eggs and egg hatching rate of schistosomiasis infected mice, causes certain damages to the tegumental structure ofschistosoma and causes morphological abnormality of germ cells, and is applicable to preparation of drugs for treating schistosomiasis.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and biomedicine, in particular to an siRNA of the SjELAV-like 1 gene of Schistosoma japonicum and its application. Background technique [0002] Schistosomiasis (Schistosomiasis) is a widely distributed and serious zoonotic parasitic disease, which is endemic; the granuloma formed in the animal liver by a large number of eggs produced by sexually mature female schistosomiasis is the root cause of host damage Reasons, and a large number of eggs excreted and caused the spread and prevalence of schistosomiasis. At present, drugs, such as praziquantel, are mainly used to treat people or animals infected with schistosomiasis, but these drugs cannot prevent reinfection of schistosomiasis. Therefore, it is necessary to find new drugs for treating schistosomiasis. In addition, antigen camouflage and antigen mimicry are important mechanisms for immune evasion of schistosomiasis, which is why th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/7105A61P33/12
CPCA61P33/12A61K31/7105C12N15/113C12N2310/14Y02A50/30
Inventor 金亚美许蓉张媛媛李肖纯程桂凤何川川李浩刘金明古少鹏
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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