Development of RNA Interference Trigger-Mediated Gene Silencing in Entamoeba invadens

Infection and Immunity
Susmitha SureshUpinder Singh

Abstract

Entamoeba histolytica, a protozoan parasite, is an important human pathogen and a leading parasitic cause of death. The organism has two life cycle stages, trophozoites, which are responsible for tissue invasion, and cysts, which are involved in pathogen transmission. Entamoeba invadens is the model system to study Entamoeba developmental biology, as high-grade regulated encystation and excystation are readily achievable. However, the lack of gene-silencing tools in E. invadens has limited the molecular studies that can be performed. Using the endogenous RNA interference (RNAi) pathway in Entamoeba, we developed an RNAi-based trigger gene-silencing approach inE. invadens We demonstrate that a gene's coding region that has abundant antisense small RNAs (sRNAs) can trigger silencing of a gene that is fused to it. The trigger fusion leads to the generation of abundant antisense sRNAs that map to the target gene, with silencing occurring independently of trigger location at the 5' or 3' end of a gene. Gene silencing is stably maintained during development, including encystation and excystation. We have used this approach to successfully silence two E. invadens genes: a putative rhomboid protease gene and a SHAQKY family Myb gene. T...Continue Reading

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Citations

May 31, 2019·Cellular Reprogramming·Han Qin, Jun Cai
Jul 15, 2020·Parasitology·Eman Abdelazeem Abuelwafa MousaFumika Mi-Ichi
Aug 9, 2018·Frontiers in Cellular and Infection Microbiology·Fumika Mi-IchiHiroki Yoshida
Nov 4, 2020·Journal of Orofacial Orthopedics = Fortschritte Der Kieferorthopädie : Organ/official Journal Deutsche Gesellschaft Für Kieferorthopädie·Jun CaiGang Yu
Dec 3, 2020·International Journal for Parasitology·Monica Mendes Kangussu-MarcolinoUpinder Singh

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