Effect of using amino acids in the freeze‑drying of siRNA lipoplexes on gene knockdown in cells after reverse transfection

  • Authors:
    • Min Tang
    • Yoshiyuki Hattori
  • View Affiliations

  • Published online on: July 12, 2021     https://doi.org/10.3892/br.2021.1448
  • Article Number: 72
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Abstract

Recently, small interfering RNA (siRNA)/cationic liposome complexes (siRNA lipoplexes) have become a crucial research tool for studying gene function. Easy and reliable siRNA transfection with a large set of siRNAs is required for the successful screening of gene function. Reverse (Rev)‑transfection with freeze‑dried siRNA lipoplexes is validated for siRNA transfection with a large set of siRNAs in a multi‑well plate. In our previous study, it was shown that Rev‑transfection with siRNA lipoplexes freeze‑dried in disaccharides or trisaccharides resulted in long‑term stability with a high level of gene‑knockdown activity. In the present study, the effects of amino acids used as cryoprotectants in the freeze‑drying of siRNA lipoplexes on gene knockdown via Rev‑transfection were assessed. A total of 15 types of amino acids were used to prepare freeze‑dried siRNA lipoplexes, and it was found that the freeze‑drying of siRNA lipoplexes with amino acid concentrations >100 mM strongly suppressed targeted gene expression regardless of the amino acid type; however, some amino acids strongly upregulated or downregulated gene expression in the cells transfected with negative control siRNA. Amongst the amino acids tested, the presence of asparagine showed specific gene‑knockdown activity, forming large cakes after freeze‑drying and retaining a favorable siRNA lipoplex size after rehydration. These findings provide valuable information regarding amino acids as cryoprotectants for Rev‑transfection using freeze‑dried siRNA lipoplexes for the efficient delivery of siRNA into cells.

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September-2021
Volume 15 Issue 3

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Spandidos Publications style
Tang M and Tang M: Effect of using amino acids in the freeze‑drying of siRNA lipoplexes on gene knockdown in cells after reverse transfection. Biomed Rep 15: 72, 2021
APA
Tang, M., & Tang, M. (2021). Effect of using amino acids in the freeze‑drying of siRNA lipoplexes on gene knockdown in cells after reverse transfection. Biomedical Reports, 15, 72. https://doi.org/10.3892/br.2021.1448
MLA
Tang, M., Hattori, Y."Effect of using amino acids in the freeze‑drying of siRNA lipoplexes on gene knockdown in cells after reverse transfection". Biomedical Reports 15.3 (2021): 72.
Chicago
Tang, M., Hattori, Y."Effect of using amino acids in the freeze‑drying of siRNA lipoplexes on gene knockdown in cells after reverse transfection". Biomedical Reports 15, no. 3 (2021): 72. https://doi.org/10.3892/br.2021.1448