The challenging nature of primary T lymphocytes for transfection: Effect of protamine sulfate on the transfection efficiency of chemical transfection reagents

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RAHIMMANESH ILNAZ; TOTONCHI MEHDI; KHANAHMAD HOSSEIN

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Journal Paper

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Journal: RESEARCH IN PHARMACEUTICAL SCIENCES Year:2020 | Volume:15 | Issue:5 Page(s): 437-446

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Title

The challenging nature of primary T lymphocytes for transfection: Effect of protamine sulfate on the transfection efficiency of chemical transfection reagents

Author(s)

RAHIMMANESH ILNAZ | TOTONCHI MEHDI | KHANAHMAD HOSSEIN | Issue Writer Certificate

Keywords

Gene transfer techniques

Protamine sulfate

T-Lymphocytes

Transfection

Abstract

Background and purpose: The optimization of an effective non-viral gene delivery method for genetic manipulation of primary human T cells has been a major challenge in immunotherapy researches. Due to the poor transfection efficiency of conventional methods in T cells, there has been an effort to increase the transfection rate in these cells. Protamine is an FDA-approved compound with a documented safety profile that enhances DNA condensation for gene delivery. Experimental approach: In this study, the effect of protamine sulfate on the transfection efficiency of standard transfection reagents, was evaluated to transfect primary human T cells. In this regard, pre-condensation of DNA was applied using protamine, and the value of the zeta potential of DNA/protamine/cargo complexes was determined. T cells were transfected with DNA/protamine/cargo complexes. The transfection efficiency rate was evaluated by flow cytometry. Also, the green fluorescent protein expression level and cytotoxicity of each complex were identified using real-time polymerase chain reaction and MTT assay, respectively. Findings/Results: Our results demonstrated that protamine efficiently increases the positive charge of DNA/cargo complex without any cytotoxic effect on the primary human T cells. We observed that the transfection efficiency in DNA/protamine/ Lipofectamine ® 2000 and DNA/protamine/TurboFect TM was 87. 2% and 78. 9%, respectively, while transfection of T cells by Lipofectamine ® 2000 and TurboFect TM would not result in sufficient transfection. Conclusion and implications: Protamine sulfate enhanced the transfection rate of T cells; and could be a promising non-viral gene delivery method to achieve a safe, rapid, cost-effective, and efficient system which will be further applied in gene therapy and T cells manipulation methods.

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APA: Copy

RAHIMMANESH, ILNAZ, TOTONCHI, MEHDI, & KHANAHMAD, HOSSEIN. (2020). The challenging nature of primary T lymphocytes for transfection: Effect of protamine sulfate on the transfection efficiency of chemical transfection reagents. RESEARCH IN PHARMACEUTICAL SCIENCES (RPS), 15(5), 437-446. SID. https://sid.ir/paper/670855/en

Vancouver: Copy

RAHIMMANESH ILNAZ, TOTONCHI MEHDI, KHANAHMAD HOSSEIN. The challenging nature of primary T lymphocytes for transfection: Effect of protamine sulfate on the transfection efficiency of chemical transfection reagents. RESEARCH IN PHARMACEUTICAL SCIENCES (RPS)[Internet]. 2020;15(5):437-446. Available from: https://sid.ir/paper/670855/en

IEEE: Copy

ILNAZ RAHIMMANESH, MEHDI TOTONCHI, and HOSSEIN KHANAHMAD, “The challenging nature of primary T lymphocytes for transfection: Effect of protamine sulfate on the transfection efficiency of chemical transfection reagents,” RESEARCH IN PHARMACEUTICAL SCIENCES (RPS), vol. 15, no. 5, pp. 437–446, 2020, [Online]. Available: https://sid.ir/paper/670855/en

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