No induction of anti-viral responses in human cell lines HeLa and MCF-7 when transfecting with siRNA or siLNA

Cecilia Dahlgren, Claes R Wahlestedt, Håkan Thonberg

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Gene silencing by RNAi and siRNAs has become a well-used tool for researchers. Because of its relatively small size, siRNA was originally thought to avoid activation of anti-viral responses. Recent reports demonstrating so-called "off-target effects" are therefore alarming. One issue raised is that siRNA induces interferon-regulated genes at the transcriptional level. We characterize the anti-viral responses of synthetic siRNA and in vitro-transcribed siRNA by measuring the mRNA levels of IFN-β and OAS2 in HeLa cells. Transfections with both traditional and LNA-modified synthetic siRNA cause no anti-viral responses, whereas transfection with either long dsRNA or in vitro-transcribed siRNA leads to greater than 1000-fold induction of these genes. The lack of response was also demonstrated at the level of phosphorylated eIF2α, and measuring of IFN-β by ELISA in cell culture media from the human cell line MCF-7. Altogether, transfection with synthetic siRNA does not induce anti-viral responses in these two cell lines. Our results reinforce the role of siRNA as an effective tool for reverse genetics and strengthen siLNA as a tool for future therapeutic applications.

Original languageEnglish
Pages (from-to)1211-1217
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume341
Issue number4
DOIs
StatePublished - Mar 24 2006
Externally publishedYes

Fingerprint

Small Interfering RNA
Cells
Cell Line
Transfection
Genes
Reverse Genetics
Virus Activation
Gene Silencing
RNA Interference
HeLa Cells
Cell culture
Interferons
Culture Media
Cell Culture Techniques
Chemical activation
Enzyme-Linked Immunosorbent Assay
Research Personnel
Messenger RNA

Keywords

  • Cell cultures
  • Interferon
  • Off-target
  • RNA interference
  • RNAi
  • siLNA
  • siRNA

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

No induction of anti-viral responses in human cell lines HeLa and MCF-7 when transfecting with siRNA or siLNA. / Dahlgren, Cecilia; Wahlestedt, Claes R; Thonberg, Håkan.

In: Biochemical and Biophysical Research Communications, Vol. 341, No. 4, 24.03.2006, p. 1211-1217.

Research output: Contribution to journalArticle

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