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Molecular characterization of RNase III protein of Asaia sp. for developing a robust RNAi-based paratransgensis tool to affect the sexual life-cycle of Plasmodium or Anopheles fitness

February 11, 2020 - 15:47 -- Open Access
Majid Asgari, Mahdokht Ilbeigikhamsehnejad, Elham Rismani, Navid Dinparast Djadid, Abbasali Raz
Parasit Vectors. 2020; 13:42

According to scientific recommendations, paratransgenesis is one of the solutions for improving the effectiveness of the Global Malaria Eradication Programme. In paratransgenesis, symbiont microorganisms are used for distorting or blocking the parasite life-cycle, affecting the fitness and longevity of vectors or reducing the vectorial competence. It has been revealed recently that bacteria could be used as potent tools for double stranded RNA production and delivery to insects. Moreover, findings showed that RNase III mutant bacteria are more competent for this aim. Asaia spp. have been introduced as potent paratransgenesis candidates for combating malaria and, based on their specific features for this goal, could be considered as effective dsRNA production and delivery tools to Anopheles spp. Therefore, we decided to characterize the rnc gene and its related protein to provide the basic required information for creating an RNase III mutant Asaia bacterium.

Gene knockdown in malaria parasites via non-canonical RNAi

January 14, 2020 - 09:31 -- Open Access
Hentzschel F, Mitesser V, Fraschka SA, Krzikalla D, Carrillo EH, Berkhout B, Bártfai R, Mueller AK, Grimm D
Nucleic Acids Research, Volume 48, Issue 1, 10 January 2020, Page e2

The lack of endogenous RNAi machinery in the malaria parasite Plasmodium hampers gene annotation and hence antimalarial drug and vaccine development. Here, we engineered rodent Plasmodium berghei to express a minimal, non-canonical RNAi machinery that solely requires Argonaute 2 (Ago2) and a modified short hairpin RNA, so-called AgoshRNA. Using this strategy, we achieved robust and specific gene knockdown throughout the entire parasite life cycle. We also successfully silenced the endogenous gene perforin-like protein 2, phenocopying a full gene knockout.

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