Journal
PLACENTA
Volume 30, Issue 2, Pages 124-129Publisher
W B SAUNDERS CO LTD
DOI: 10.1016/j.placenta.2008.10.003
Keywords
BeWo; Syncytiotrophoblast cells; First trimester placenta; Transfection; Placental alkaline phosphatase; siRNA
Funding
- BBSRC
- Wellcome Trust
- Biotechnology and Biological Sciences Research Council [BB/E007678/1] Funding Source: researchfish
- BBSRC [BB/E007678/1] Funding Source: UKRI
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The use of RNA interference (RNAi) to deplete individual proteins from cells or tissue has revolutionised our ability to characterise gene function. The placenta is an attractive target for studies in which the role of specific proteins can be compared with cell culture models and explanted villous tissue where physiological function can be maintained ex vivo. In this study, we compared a variety of commercially available reagents and approaches to define methods for efficient delivery of siRNA to placental cells. Protocols optimised using fluorescently-labelled siRNA were Subsequently tested using siRNA sequences that target placental alkaline phosphatase (PLAP), chosen because of its high abundance in trophoblast. mRNA abundance was assayed using qRT-PCR, and the effect on protein was examined using immunolocalisation. We report that different protocols are required for BeWo choriocarcinoma cells (nucleofection), primary cytotrophoblast cells (lipid-based transfection) and villous tissue explants (nucleofection). The results provide guidelines for optimal siRNA-mediated knockdown in these three models of the human placenta. (C) 2008 Elsevier Ltd. All rights reserved.
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