High-efficiency siRNA-based gene knockdown in human embryonic stem cells

  1. Caihong Qiu1
  1. 1Yale Stem Cell Center, New Haven, Connecticut 06520, USA
  2. 2Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, New Haven, Connecticut 06510, USA
  3. 3Wenzhou University College of Teacher Education, Wenzhou, Zhejiang 325035, China

Abstract

Loss-of-function studies in human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) via nonviral approaches have been largely unsuccessful. Here we report a simple and cost-effective method for high-efficiency delivery of plasmids and siRNAs into hESCs and iPSCs. Using this method for siRNA delivery, we achieve >90% reduction in the expression of the stem cell factors Oct4 and Lin28, and observe cell morphological and staining pattern changes, characteristics of hESC differentiation, as a result of Oct4 knockdown.

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Footnotes

  • Reprint requests to: Yingqun Huang, Yale Stem Cell Center, PO Box 208073, New Haven, CT 06520, USA, or Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, New Haven, CT 06510, USA; e-mail: yingqun.huang{at}yale.edu; fax: (203) 785-7134; or Caihong Qiu, Yale Stem Cell Center, PO Box 208073, New Haven, CT 06520, USA; e-mail: caihong.qiu{at}yale.edu; fax: (203) 785-4305.

  • Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.2350710.

  • Received July 4, 2010.
  • Accepted September 16, 2010.
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