Book chapter
Small Interfering RNA (siRNA) Transfection in Epiblast Stem Cells
Epiblast Stem Cells: Methods and Protocols, pp.47-55
Methods in Molecular Biology, 2490, Humana New York
2022
PMID: 35486238
Abstract
Lipid-based transfection of siRNA is a technique routinely used to investigate gene function in experiments using mammalian cells cultured in vitro. Due to innate differences in cellular characteristics, the efficiency of lipid-based transfection is variable across cell types. Pluripotent cells which exist in a "primed" state such as human embryonic stem cells (hESCs) and mouse epiblast stem cells (mEpiSCs) are notorious for being refractory to lipid-based transfection systems. Herein we describe a forward transfection protocol which we routinely use to achieve upwards of 70% transfection efficiency rates in mEpiSCs. Our protocol also includes a suggested transfection timeline and details pertaining to the techniques we use to validate transfection success.
Details
- Title
- Small Interfering RNA (siRNA) Transfection in Epiblast Stem Cells
- Authors
- Georgia R Kafer (Author) - University of the Sunshine Coast, Queensland, School of Health and Behavioural Sciences - Legacy
- Contributors
- Pierre Osteil (Editor) - École Polytechnique Fédérale de Lausanne
- Publication details
- Epiblast Stem Cells: Methods and Protocols, pp.47-55
- Series
- Methods in Molecular Biology; 2490
- Publisher
- Humana New York
- Date published
- 2022
- DOI
- 10.1007/978-1-0716-2281-0_5; 10.1007/978-1-0716-2281-0
- ISSN
- 1940-6029
- PMID
- 35486238
- Organisation Unit
- School of Health - Biomedicine; Australian Centre for Pacific Islands Research; University of the Sunshine Coast, Queensland; School of Health and Sport Sciences - Legacy; Centre for Bioinnovation; School of Health and Behavioural Sciences - Legacy
- Language
- English
- Record Identifier
- 99634540702621
- Output Type
- Book chapter
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