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Double-stranded RNA Mediates Selective Gene Silencing of Protein Phosphatase Type 1 Delta Isoform in HEK-293 Cells |
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| Authors: | Hiroyuki Morimoto Hirohiko Okamura Kaya Yoshida Seiichiro Kitamura Tatsuji Haneji |
| Institution: | 1. Department of Anatomy, School of Dentistry, The University of Tokushima, Kuramoto, Tokushima, 770-8504, Japanmorimoto@dent.tokusnima-u.ac.jp;3. Department of Histology and Oral Histology, School of Dentistry, The University of Tokushima, Kuramoto, Tokushima, 770-8504, Japan;4. Department of Anatomy, School of Dentistry, The University of Tokushima, Kuramoto, Tokushima, 770-8504, Japan |
| 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 PP18. 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. |
| Keywords: | Protein phosphatases Isoform Double-stranded RNA RNAi HEK-293 cells |