Use of PepFect14 and siRNA for Targeted Gene Silencing in Human Embryonic Stem Cells.

Egle-Helene Ervin, Margus Pooga, Toivo Maimets

Methods in molecular biology (Clifton, N.J.) 2022

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The unique properties of human embryonic stem (hES) cells render them invaluable for many scientific and clinical endeavors. Wider application of hES cells requires a comprehensive understanding of their biology that can be dissected using RNA interference-based gene silencing. However, commonly used transfection methods to deliver nucleic acids into a cell often lead to differentiation of hES cells. For that reason, effective transfection technique with minimal side effects is essential for studying and employing hES cells. Here, we describe a CPP-based method for targeted gene silencing in hES cells using siRNA complexed with PepFect 14 (PF14). This approach results in effective downregulation of mRNA and protein levels of a target gene without adverse effects on cell viability and pluripotency. © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Egle-Helene Ervin, Margus Pooga, Toivo Maimets. Use of PepFect14 and siRNA for Targeted Gene Silencing in Human Embryonic Stem Cells. Methods in molecular biology (Clifton, N.J.). 2022;2383:579-585