Small Interfering RNA (siRNA) Transfection in Epiblast Stem Cells.
Epiblast
Lipid
Lipofectamine
Mouse
Small interfering RNA
Transfection
mEpiSC
siRNA
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
29
4
2022
pubmed:
30
4
2022
medline:
4
5
2022
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 35486238
doi: 10.1007/978-1-0716-2281-0_5
doi:
Substances chimiques
Lipids
0
RNA, Small Interfering
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
47-55Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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