Transient and stable transfection in the protozoan parasite Entamoeba invadens

Entamoeba histolytica is an important human pathogen and a major health problem worldwide. Many aspects of parasite biology can be studied with the exception of stage conversion, which cannot be reproduced adequately in E. histolytica. The reptile parasite Entamoeba invadens is a vital model system for studying stage conversion since it can be induced to undergo both encystation and excystation with high efficiency in vitro. However, functional studies using E. invadens have been limited by the lack of genetic tools in this species. Here, we report a new method for both transient and stable transfection of E. invadens. These new tools will greatly enhance research into Entamoeba development.

For stable transfection of Entamoeba invadens we utilized neomycin drug selection. RT-PCR demonstrates expression of the EiMyb gene in the stable transfectant cell line, but not in WT (untransfected) parasites.Figure optionsDownload high-quality image (80 K)Download as PowerPoint slideHighlights
► Entamoeba invadens is a model system for studying Entamoeba stage conversion.
► Lack of genetic tools has rendered this system less useful for molecular studies.
► We have developed methods for transient and stable transfection of E. invadens.
► These will be valuable tools for studying molecular basis of parasite development.