A simple and innovative method for cell membrane passing the small interfering RNA (siRNA) in eukaryotic cells; toxoplasma gondii as an experience

Abstract

Background: Eukaryotic cells have membrane organelles. Due to the high prevalence of toxoplasmosis and lack of effective drugs especially on dormant cystic stages, we aimed adding small interfering RNA (siRNA) in culture medium on the proliferation period of microorganism in order to transmit siRNA into the tachyzoite of Toxoplasma gondii (T. gondii) named addition before barrier arise style (ABBAS). Methods: T. gondii was proliferated in mouse. Based on dihydrofolate reductase gene sequence, was designed siRNA using in-silico method. The flow cytometry method was used to estimate the transmission rate of tagged siRNA into the parasite. Statistical analysis was accomplished using SPSS software. Findings: The transmission rate of tagged siRNA into the parasite was 73 using ABBAS. The mean death time of siRNA-treated mice was approximately 2 days later than control group. Conclusion: T. gondii possess an inner membrane complex (IMC). Probably success of this method in passing more siRNA into the cytoplasm of tachyzoites is due to the fact that siRNA molecule enters the cell prior to absolute formation of the membrane or when the membrane possesses larger pores. © 2017, Isfahan University of Medical Sciences(IUMS). All rights reserved.