Double-stranded RNA mediates selective gene silencing of protein phosphatase type 1 delta isoform in HEK-293 cells

Hiroyuki Morimoto, Hirohiko Okamura, Kaya Yoshida, Seiichiro Kitamura, Tatsuji Haneji

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The reversible phosphorylation of proteins mediates cellular signals in eukaryotic cells. RNA interference inhibits the expression of genes and proteins in a sequence-specific manner and provides a tool to study the functions of target molecules. The effect of RNA interference on protein phosphatase isoforms in HEK-293 cells was examined. Protein phosphatase 1 delta (PP1δ) sequence-specific double-stranded RNA (dsRNA) inhibited mRNA and protein expression of the PP1δ. This RNA interference did not affect the expression of α and γ1 isoforms of PP1. Transfection of antisense RNA specific for PP1δ also suppressed the expression of PP1δ. It was further demonstrated by an in vitro RNA cleavage assay that extracts of HEK-293 cells catalyzed the processing of dsRNA. This cell line had much stronger mRNA expression of Dicer, an RNase III-like enzyme, than did human osteoblastic MG63 cells. The present results show that RNA interference is a useful tool to distinguish between PP1 isoforms.

Original languageEnglish
Pages (from-to)327-331
Number of pages5
JournalJournal of Enzyme Inhibition and Medicinal Chemistry
Volume19
Issue number4
DOIs
Publication statusPublished - Aug 2004
Externally publishedYes

Keywords

  • Double-stranded RNA
  • HEK-293 cells
  • Isoform
  • Protein phosphatases
  • RNAi

ASJC Scopus subject areas

  • Pharmacology
  • Drug Discovery

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