siRNA of schistosoma japonicum katsurada SjLwr gene 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 drug resistance, affecting clinical application effects, and large first-pass effects

Pending Publication Date: 2019-12-13
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

At present, drugs such as praziquantel are mainly used in the treatment of schistosomiasis, but when praziquantel is taken orally, the "first-pass effe

Method used

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  • siRNA of schistosoma japonicum katsurada SjLwr gene and application thereof
  • siRNA of schistosoma japonicum katsurada SjLwr gene and application thereof
  • siRNA of schistosoma japonicum katsurada SjLwr gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: In vivo screening of small RNAs

[0028] 1. Method steps

[0029] 1.1 siRNA molecules and wxya Preparation of primers for quantitative real-time PCR of genes

[0030] Find schistosomes on the NCBI website (http: / / www.ncbi.nlm.nih.gov / ) Lwr Gene sequence (FN321279.1). according to Lwr Expression of gene open reading frame in Schistosoma japonicum, design and synthesis of three pairs of dsRNA to produce Lwr Gene-specific inhibitory effects, affecting Lwr gene expression. Simultaneously, a pair of unrelated control dsRNA (NCsiRNA) was synthesized.

[0031] S1 dsRNA: sense: 5'-GCGUUAUUCAAUGCCGCAUTT-3'(SEQ NO.1)

[0032] anti-sence: 5'-AUGCGGCAUUGAAUAACGCTT-3'(SEQ NO.2)

[0033] S2 dsRNA: sense:5'-GCUACAGUCCAACCACAAATT-3'(SEQ NO.3)

[0034] anti-sence: 5'-UUUGUGGUUGGACUGUAGGTT-3'(SEQ NO.4)

[0035]S3 dsRNA: sense:5'-GCUACUUGACUCCCUACUUTT-3'(SEQ NO.5)

[0036] anti-sence:5'-AAGUAGGGAGUCAAGUAGCTT-3'(SEQ NO.6)

[0037] NC dsRNA:sense:5'-UUCUCCGAACG...

Embodiment 2

[0050] Embodiment 2: RNA interference experiment

[0051] 1. Method steps

[0052] 1.1 dsRNA injection

[0053] The experiment consisted of control group, unrelated control group (NC group) and S2 dsRNA treatment group, with three mice in each group. 11 days, 15 days, 19 days, 23 days, 27 days, 31 days, 35 days and 39 days after infection, 200ul DEPC water, 200ul NC dsRNA (8µg) and 200ul SjLwr gene S2 dsRNA (8ug ), a total of 8 times.

[0054] 1.2 Hatch rate of liver worm eggs

[0055] After dissecting the mouse, take the liver and weigh it, add dechlorinated water to make it up to 20 ml, mix it with a homogenizer, take 4 ml of the crushed liquid and add it to the chicken heart bottle (add dechlorinated water to the neck of the chicken heart bottle 3-5 cm above), put cotton on the liquid surface of the chicken heart bottle, add 8 ml of dechlorinated water to the cotton, incubate at 26-27 ℃ for 2 h, transfer 8 ml of the upper layer of cotton to a 15 ml conical centrifuge A...

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Abstract

The invention discloses siRNA of a schistosoma japonicum SjLwr gene. The siRNA has nucleotide sequences as shown in SEQ ID NO.1 and SEQ ID NO.2; or nucleotide sequences shown in SEQ ID NO.3 and SEQ IDNO.4, or nucleotide sequences shown in SEQ ID NO. 5 and SEQ ID NO. 6. The invention also discloses an application of the siRNA of the schistosoma japonicum SjLwr gene. The siRNA of the schistosoma japonicum katsurada SjLwr gene can obviously inhibit transcription of SjLwr gene, can obviously reduce schistosome infected mouse insect body number, liver egg number, and liver egg contribution rate and egg hatching rate of each female insect, has certain pathological influence on insect integument structure and gonad cell morphology, and is suitable for preparation of drugs for prevention and treatment of schistosomiasis.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and biomedicine, in particular to a Schistosoma japonicum wxya Gene siRNA and its application. Background technique [0002] Schistosomiasis is a zoonotic parasitic disease with a wide range of epidemics and causes serious pathological damage to the host. At present, drugs such as praziquantel are mainly used for the treatment of schistosomiasis, but when praziquantel is taken orally, the "first-pass effect" is large and the bioavailability is poor, which affects the clinical application effect. Resistance develops and cannot be used to treat reinfection. Therefore, it is necessary to find new drugs for the treatment of schistosomiasis. [0003] Lesswright ( lwr ), a UBC9 homologue, mediates dissociation of heterochromatin regions at the end of meiotic prophase I in Drosophila melanogaster. Lwr It also has important implications for the regulation of primordial germ cells and th...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K31/7088A61P33/12
CPCC12N15/113A61K31/7088A61P33/12C12N2310/14Y02A50/30
Inventor 金亚美李肖纯程贵凤秦芳林刘金明李浩
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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