Stability and cellular uptake of polymerized siRNA (poly-siRNA)/polyethylenimine (PEI) complexes for efficient gene silencing

Seung Young Lee, Myung Sook Huh, Seulki Lee, So Jin Lee, Hyunjin Chung, Jae Hyung Park, Yu Kyoung Oh, Kuiwon Choi, Kwangmeyung Kim, Ick Chan Kwon

Research output: Contribution to journalArticle

Abstract

Small interfering RNA (siRNA) is a promising biological strategy for treatment of diverse diseases, but the therapeutic application of siRNA has been limited by its instability and poor cellular uptake efficiency. Although the development of various gene delivery systems has increased the siRNA delivery efficiency, many problems still remain to be resolved before the clinical application of siRNA. In this study, we suggest reducible polymerized siRNA a possible solution for low delivery efficiency of siRNA. Dithiol-modified red fluorescent protein (RFP) siRNAs at the 5'-ends of both sense and anti-sense strands were disulfide-polymerized. Polymerized siRNA (poly-siRNA) was composed of 30% oligomeric siRNA (50~300. bps) and 66% polymeric siRNA (above ~ 300. bps) as fractions, and was reducible in reducing solution through disulfide bond cleavage. Poly-siRNA formed more condensed and nano-sized complexes with low molecular weight polyethylenimine (PEI) by strong electrostatic interaction based on the higher charge density of poly-siRNA, compared with siRNA (mono-siRNA). The compact poly-siRNA/PEI complexes prevented the loss and degradation of siRNA from a polyanion competitor and RNases in serum. Furthermore, poly-siRNA/PEI complexes exhibited superior intracellular uptake by murine melanoma cells (B16F10), and was accompanied with RFP gene silencing efficiency of about 80%, compared to untreated cells. These results sufficiently support that strong polyanionic and reducible poly-siRNA can be utilized as a novel powerful therapeutic strategy for human diseases.

Original languageEnglish (US)
Pages (from-to)339-346
Number of pages8
JournalJournal of Controlled Release
Volume141
Issue number3
DOIs
StatePublished - Feb 2010
Externally publishedYes

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Polyethyleneimine
Gene Silencing
Small Interfering RNA
Disulfides
Gene Transfer Techniques

Keywords

  • Electrostatic complexes
  • Gene silencing
  • Polyethylenimine
  • Polymerized siRNA
  • SiRNA

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

Stability and cellular uptake of polymerized siRNA (poly-siRNA)/polyethylenimine (PEI) complexes for efficient gene silencing. / Lee, Seung Young; Huh, Myung Sook; Lee, Seulki; Lee, So Jin; Chung, Hyunjin; Park, Jae Hyung; Oh, Yu Kyoung; Choi, Kuiwon; Kim, Kwangmeyung; Kwon, Ick Chan.

In: Journal of Controlled Release, Vol. 141, No. 3, 02.2010, p. 339-346.

Research output: Contribution to journalArticle

Lee, Seung Young ; Huh, Myung Sook ; Lee, Seulki ; Lee, So Jin ; Chung, Hyunjin ; Park, Jae Hyung ; Oh, Yu Kyoung ; Choi, Kuiwon ; Kim, Kwangmeyung ; Kwon, Ick Chan. / Stability and cellular uptake of polymerized siRNA (poly-siRNA)/polyethylenimine (PEI) complexes for efficient gene silencing. In: Journal of Controlled Release. 2010 ; Vol. 141, No. 3. pp. 339-346.
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abstract = "Small interfering RNA (siRNA) is a promising biological strategy for treatment of diverse diseases, but the therapeutic application of siRNA has been limited by its instability and poor cellular uptake efficiency. Although the development of various gene delivery systems has increased the siRNA delivery efficiency, many problems still remain to be resolved before the clinical application of siRNA. In this study, we suggest reducible polymerized siRNA a possible solution for low delivery efficiency of siRNA. Dithiol-modified red fluorescent protein (RFP) siRNAs at the 5'-ends of both sense and anti-sense strands were disulfide-polymerized. Polymerized siRNA (poly-siRNA) was composed of 30{\%} oligomeric siRNA (50~300. bps) and 66{\%} polymeric siRNA (above ~ 300. bps) as fractions, and was reducible in reducing solution through disulfide bond cleavage. Poly-siRNA formed more condensed and nano-sized complexes with low molecular weight polyethylenimine (PEI) by strong electrostatic interaction based on the higher charge density of poly-siRNA, compared with siRNA (mono-siRNA). The compact poly-siRNA/PEI complexes prevented the loss and degradation of siRNA from a polyanion competitor and RNases in serum. Furthermore, poly-siRNA/PEI complexes exhibited superior intracellular uptake by murine melanoma cells (B16F10), and was accompanied with RFP gene silencing efficiency of about 80{\%}, compared to untreated cells. These results sufficiently support that strong polyanionic and reducible poly-siRNA can be utilized as a novel powerful therapeutic strategy for human diseases.",
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AU - Lee, So Jin

AU - Chung, Hyunjin

AU - Park, Jae Hyung

AU - Oh, Yu Kyoung

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