Method of transfecting interfering RNA to Echinococcus granulosus by electroporation

Abstract

The invention discloses a method for transfecting an interfering RNA to echinococcus granulosus by virtue of electroporation. The method comprises the following steps: (1) cultivating echinococcus granulosus protoscolex in an RPMI1640 culture medium containing penicillin and streptomycin for 1-5 days; (2) cleaning the cultivated echinococcus granulosus protoscolex, adding the cleaned echinococcus granulosus protoscolex to the RPMI1640 culture medium, adding transfer buffer and an interfering RNA to an electrode cup, and transfecting the echinococcus granulosus protoscolex by virtue of an electric transfer instrument; (3) standing the transfected echinococcus granulosus protoscolex and then transferring to a sterile culture dish containing the RPMI1640 culture medium, and cultivating in an incubator; and (4) detecting the GRP78mRNA expression level in the transfected echinococcus granulosus protoscolex through RT-PCR, and detecting the GRP78 protein expression level in the transfected echinococcus granulosus protoscolex through Western blot, and identifying the transfection effect. The observation result shows that the interfering RNA is successfully transfected to the echinococcus granulosus by using an electroporation method.

Description

technical field [0001] The invention discloses a method for transfecting interference RNA to Echinococcus granulosus by using electroporation, and belongs to the technical field of biological experiments. Background technique [0002] Echinococcosis (echinococcosis), commonly known as hydatid disease, is a zoonotic disease that seriously endangers human health and the development of animal husbandry caused by the middle-stage larvae of the echinococcosis tapeworm parasitizing in humans and some other animals. suffering from parasitic diseases. There are many types of echinococcus, currently in my country there are two main types: cystic echinococcosis (cystic echinococcosis, CE) and alveolar echinococcosis (AE). , which are caused by Echinococcus granulosus (E.g) and Echinococcus multilocularis (E.m) parasitizing in humans and some animals respectively. Echinococcus granulosus is distributed globally and is mainly prevalent in Xinjiang, Qinghai, Inner Mongolia, Tibet and ot...

AI Technical Summary

Problems solved by technology

For Echinococcus granulosus, how to introduce small molecule interfering RNA into the target body is a technical problem in this field. Common methods such as soaking method are not ideal, and transfection methods, such as virus-mediated method, Adenovirus, cationic liposome method, etc., research and development found that transfection of Echinococcus granulosus protoscole by liposome method and adenovirus method did not achieve the purpose of interfering RNA silencing the target gene of the protoscole. There is an urgent need for a transfection method that is simple to operate, has clear conditions, and can effectively realize the function of interfering RNA

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