The modification of siRNA with 3' cholesterol to increase nuclease protection and suppression of native mRNA by select siRNA polyplexes

Vishakha V. Ambardekar, Huai Yun Han, Michelle L. Varney, Serguei V. Vinogradov, Rakesh K Singh, Joseph A Vetro

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Polymer-siRNA complexes (siRNA polyplexes) are being actively developed to improve the therapeutic application of siRNA. A major limitation for many siRNA polyplexes, however, is insufficient mRNA suppression. Given that modifying the sense strand of siRNA with 3' cholesterol (chol-siRNA) increases the activity of free nuclease-resistant siRNA in vitro and in vivo, we hypothesized that complexation of chol-siRNA can increase mRNA suppression by siRNA polyplexes. In this study, the characteristics and siRNA activity of self assembled polyplexes formed with chol-siRNA or unmodified siRNA were compared using three types of conventional, positively charged polymers: (i) biodegradable, cross-linked nanogels (BDNG) (ii) graft copolymers (PEI-PEG), and (iii) linear block copolymers (PLL10-PEG, and PLL50-PEG). Chol-siRNA did not alter complex formation or the resistance of polyplexes to siRNA displacement by heparin but increased nuclease protection by BDNG, PLL10-PEG, and PLL50-PEG polyplexes over polyplexes with unmodified siRNA. Chol-CYPB siRNA increased suppression of native CYPB mRNA in mammary microvascular endothelial cells (MVEC) by BDNG polyplexes (35%) and PLL10-PEG polyplexes (69%) over comparable CYPB siRNA polyplexes but had no effect on PEI-PEG or PLL50-PEG polyplexes. Overall, these results indicate that complexation of chol-siRNA increases nuclease protection and mRNA suppression by select siRNA polyplexes. These results also suggest that polycationic block length is an important factor in increasing mRNA suppression by PLL-PEG chol-siRNA polyplexes in mammary MVEC.

Original languageEnglish (US)
Pages (from-to)1404-1411
Number of pages8
JournalBiomaterials
Volume32
Issue number5
DOIs
StatePublished - Feb 1 2011

Fingerprint

Cholesterol
Small Interfering RNA
Polyethylene glycols
Messenger RNA
Polyetherimides
Endothelial cells
Complexation
Biodegradable polymers
Graft copolymers
Phase locked loops
Block copolymers
Polymers
Breast
Endothelial Cells
Cross Protection

Keywords

  • H-2Kb-ts-A58
  • Immortomouse
  • Mammary fat pad
  • PEGss
  • PEGylated cationic polymers
  • PPIB

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Cite this

The modification of siRNA with 3' cholesterol to increase nuclease protection and suppression of native mRNA by select siRNA polyplexes. / Ambardekar, Vishakha V.; Han, Huai Yun; Varney, Michelle L.; Vinogradov, Serguei V.; Singh, Rakesh K; Vetro, Joseph A.

In: Biomaterials, Vol. 32, No. 5, 01.02.2011, p. 1404-1411.

Research output: Contribution to journalArticle

Ambardekar, Vishakha V. ; Han, Huai Yun ; Varney, Michelle L. ; Vinogradov, Serguei V. ; Singh, Rakesh K ; Vetro, Joseph A. / The modification of siRNA with 3' cholesterol to increase nuclease protection and suppression of native mRNA by select siRNA polyplexes. In: Biomaterials. 2011 ; Vol. 32, No. 5. pp. 1404-1411.
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