Silencing genes by RNA interference in the protozoan parasite Entamoeba histolytica.

Experimental procedures using the RNA interference (RNAi) approach have recently emerged as a powerful tool for gene silencing in eukaryotic microbes for which gene replacement techniques have not yet been developed. Our group has recently explored RNAi to knock down gene-specific expression in the protozoan parasite Entamoeba histolytica, through delivery of small interfering RNA (siRNA) oligonucleotides by the soaking approach. Standardized conditions for the soaking of E. histolytica trophozoites with siRNAs result in highly specific and significant silencing of parasite cognate genes. Real-time PCR analysis indicates that a 16-hour treatment with siRNAs usually results in half-extinction of target messenger RNA. Furthermore, Western blot analysis of trophozoite crude extracts with the use of specific antibodies shows a similar reduction of cognate protein levels after siRNA treatment.