Cellular siRNA delivery mediated by a cell-permeant RNA-binding protein and photoinduced RNA interference

Tamaki Endoh, Masahiko Sisido, Takashi Ohtsuki

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

HIV-1 TAT peptide, which is a cell-penetrating peptide (CPP), was fused to the U1A RNA-binding domain (TatU1A) to generate a sequence-specific siRNA delivery system for mammalian cells. The siRNA contained a short 5′-extension that is specifically recognized by the U1A RNA-binding domain (U1AsiRNA). Specific binding of TatU1A to the U1AsiRNA was confirmed using a gel mobility shift assay. The U1AsiRNA was internalized by cells only when it was preincubated with TatU1A before addition to the cells. Although most of the internalized siRNA seemed to be entrapped in endocytic compartments, efficient redistribution of the entrapped siRNAs was achieved by photostimulation of a fluorophore attached to TatU1A. Once in the cytoplasm, the siRNA induced RNAi-mediated gene silencing. We refer to this delivery strategy as CLIP-RNAi. CLIP-RNAi is a promising strategy for RNAi experiments and for pinpoint RNAi therapy.

Original languageEnglish
Pages (from-to)1017-1024
Number of pages8
JournalBioconjugate Chemistry
Volume19
Issue number5
DOIs
Publication statusPublished - May 2008

Fingerprint

RNA-Binding Proteins
RNA Interference
RNA
Peptides
Small Interfering RNA
Fluorophores
Assays
Gels
Genes
Cells
Cell-Penetrating Peptides
Gene Silencing
Electrophoretic Mobility Shift Assay
HIV-1
Cytoplasm
Experiments
Carrier Proteins
RNA-Binding Motifs

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Cellular siRNA delivery mediated by a cell-permeant RNA-binding protein and photoinduced RNA interference. / Endoh, Tamaki; Sisido, Masahiko; Ohtsuki, Takashi.

In: Bioconjugate Chemistry, Vol. 19, No. 5, 05.2008, p. 1017-1024.

Research output: Contribution to journalArticle

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