Small interfering ribonucleic acid (siRNA) of schistosoma japonicum katsurada membrane-associated progesterone receptor component (PGMRC2) gene and application thereof

A technology for schistosomiasis and schistosomiasis, applied in the fields of molecular biology and biomedicine, can solve serious problems, the prospect of schistosomiasis is not optimistic, and there are no drugs for schistosomiasis, etc.

Inactive Publication Date: 2014-03-26
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although my country has made great achievements in the prevention and control of schistosomiasis, the prospect of comprehensive control of schistosomiasis transmission is still not optimistic due to the difficulty of eradicating the intermediate host snails and the serious phenomenon of repeated infection of schistosomia...

Method used

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  • Small interfering ribonucleic acid (siRNA) of schistosoma japonicum katsurada membrane-associated progesterone receptor component (PGMRC2) gene and application thereof
  • Small interfering ribonucleic acid (siRNA) of schistosoma japonicum katsurada membrane-associated progesterone receptor component (PGMRC2) gene and application thereof
  • Small interfering ribonucleic acid (siRNA) of schistosoma japonicum katsurada membrane-associated progesterone receptor component (PGMRC2) gene and application thereof

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Experimental program
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Effect test

Embodiment 1

[0023] Example 1 In vitro RNA interference

[0024] 1. Design of siRNA molecules

[0025] Four pairs of siRNA molecules were designed according to the PGMRC2 gene sequence of Schistosoma japonicum (GeneBank: FN317689.1) (see Table 1 below). The NC-RL siRNA molecule was an irrelevant control, and the siRNA was designed and synthesized by Shanghai Jima Pharmaceutical Technology Co., Ltd.

[0026] Table 1 siRNA sequence

[0027]

[0028] 1.2 Collection of worms

[0029] The abdominal patch was used to attack and infect New Zealand white rabbits, each infected with about 5000 Schistosoma japonicum cercariae. After the 14th day of infection, the worms collected by the hepatic vein perfusion method were dissected and washed once quickly with warm PBS containing double antibodies, and then washed three times with worm culture medium in an ultra-clean table.

[0030] 1.3 Processing of rabbit red blood cells

[0031] ①Suck 0.1ml heparin from the syringe, collect 1ml blood from the ear vein of the...

Embodiment 2

[0038] Example 2 RNA interference in vivo

[0039] 1. Method steps

[0040] BALB / c mice were divided into three groups: blank control group, irrelevant interference control group and siRNA treatment group, with five mice in each group. The abdominal patch was used to infect cercariae of Schistosoma japonicum, and each mouse infected about 100. Beginning on the 13th day after infection, the mice in the blank control group were injected with 0.1ml PBS solution (pH=7.4) per day in the tail vein, and the mice in the control group were injected with 0.1ml NC-RL(50ug) / PBS(pH=7.4) per day in the tail vein. =7.4) solution. Mice in the siRNA treatment group were injected with 0.1 ml of SjPGMRC2 gene siRNA (50ug) / PBS (pH=7.4) solution every day via tail vein. After 7 consecutive injections, the worms were dissected and the hepatic portal vein was used to flush the worms. Observe and count the worms. Same as above, using Real-time PCR to detect changes in the transcription level of PGMRC2 ...

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Abstract

The invention discloses a small interfering ribonucleic acid (siRNA) of a schistosoma japonicum katsurada membrane-associated progesterone receptor component (PGMRC2) gene. The siRNA is selected from a pair or combination of any more than two pairs of nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2, nucleotide sequences shown in SEQ ID NO.3 and SEQ ID NO.4, nucleotide sequences shown in SEQ ID NO.5 and SEQ ID NO.6, and nucleotide sequences shown in SEQ ID NO.7 and SEQ ID NO.8. The invention also discloses application of the siRNA of the schistosoma japonicum katsurada PGMRC2 gene. By adopting the siRNA of the schistosoma japonicum katsurada PGMRC2 gene disclosed by the invention, transcription of the Sj PGMRC2 gene can be obviously restricted, and 24% of worm reduction rate can be obtained. Therefore, the siRNA of the schistosoma japonicum katsurada PGMRC2 gene can be applicable to preparation of a medicament for treating bilharziasis.

Description

Technical field [0001] The present invention relates to the technical fields of molecular biology and biomedicine, in particular to a siRNA of Schistosoma japonicum PGMRC2 (membrane-associated progesterone receptor component 2) gene and its application. Background technique [0002] Schistosomiasis japonica is an important zoonotic parasitic disease. Although my country has made great achievements in the prevention and control of schistosomiasis, the prospect of comprehensive control of the spread of schistosomiasis is still not optimistic due to the difficulty of eliminating the intermediate host oncomelania and the serious phenomenon of repeated schistosomiasis infection. Since its inception, praziquantel has been widely used in the clinical treatment and prevention of schistosomiasis due to its high efficiency, low toxicity, convenient use, and low price. It has become the only drug currently used for the treatment of schistosomiasis. It has not been seen for more than 20 year...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P33/12
Inventor 傅志强吴秀娟林矫矫洪炀石耀军陆珂
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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