Polyethylenimine-cationized β-catenin protein transduction activates the Wnt canonical signaling pathway more effectively than cationic lipid-based transduction

Midori Kitazoe, Junichiro Futami, Mitsuo Nishikawa, Hidenori Yamada, Yoshitake Maeda

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The Wnt canonical signaling pathway is essential for the early development of eukaryotic organisms and plays a key role in cell proliferation, differentiation, and oncogenesis. Moreover, the Wnt canonical signaling pathway contributes to the self-renewal of mouse hematopoietic stem cells (HSCs). Here, we demonstrate artificial activation of the Wnt canonical signaling pathway by β-catenin protein transduction. Constitutively active β-catenin protein was introduced into human embryonic kidney HEK-293 cells using a polyethylenimine (PEI) cationization method, or with the BioPORTER protein transduction reagent. We have previously shown that modification with PEI effectively causes proteins to be internalized by living mammalian cells. PEI-cationized, constitutively active β-catenin protein was added to HEK-293 cells, and induction of several Wnt/β-catenin target genes was detected by real-time PCR. However, using BioPORTER to introduce active β-catenin did not activate the Wnt canonical signaling pathway. Introduction of eGFPNuc (enhanced green fluorescent protein variant containing a nuclear localization signal) into HEK-293 cells using the BioPORTER reagent caused significant cell death, as determined by propidium iodide staining. In contrast, the PEI-modified eGFPNuc did not impair survival of HEK-293 cells. These results indicate that the Wnt canonical signaling pathway could be successfully activated by transduction of PEI-cationized active β-catenin, and the PEI-cationization method is an effective and safe technology for protein transduction into mammalian cells.

Original languageEnglish
Pages (from-to)385-392
Number of pages8
JournalBiotechnology Journal
Volume5
Issue number4
DOIs
Publication statusPublished - Apr 2010

Fingerprint

Polyethyleneimine
Catenins
Wnt Signaling Pathway
HEK293 Cells
Lipids
Nuclear Localization Signals
Proteins
Propidium
Hematopoietic Stem Cells
Real-Time Polymerase Chain Reaction
Cell Differentiation
Carcinogenesis
Cell Death
Cell Proliferation
Staining and Labeling
Technology
Kidney
Genes

Keywords

  • BioPORTER
  • Polyethylenimine (PEI)
  • Protein transduction
  • Protein transduction domain (PTD)
  • Wnt canonical signaling pathway

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

Cite this

Polyethylenimine-cationized β-catenin protein transduction activates the Wnt canonical signaling pathway more effectively than cationic lipid-based transduction. / Kitazoe, Midori; Futami, Junichiro; Nishikawa, Mitsuo; Yamada, Hidenori; Maeda, Yoshitake.

In: Biotechnology Journal, Vol. 5, No. 4, 04.2010, p. 385-392.

Research output: Contribution to journalArticle

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