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. 2022:2383:579-585.
doi: 10.1007/978-1-0716-1752-6_37.

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

Egle-Helene Ervin  1   2 , Margus Pooga  3 , Toivo Maimets  4
Affiliations

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

Egle-Helene Ervin et al. Methods Mol Biol. 2022.

Abstract

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.

Keywords: B2M; Human embryonic stem cells; OCT4; PepFect 14; Transfection; siRNA.

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