A Simple and Innovative Method for Cell Membrane Passing the Small Interfering RNA (siRNA) in Eukaryotic Cells; Toxoplasma Gondii as an Experience

Document Type : Original Article (s)

Authors

1 Associate Professor, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 MSc Student, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

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 SPSSsoftware.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.

Keywords

References

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Journal of Isfahan Medical School
Volume 35, Issue 431 - Serial Number 431
March and April 2017
Pages 571-576

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History

  • Receive Date: 28 January 2017
  • Revise Date: 23 May 2024
  • Accept Date: 06 April 2022

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