Enhanced delivery of siRNA complexes by sonoporation in transgenic rice cell suspension cultures

J Microbiol Biotechnol. 2009 Aug;19(8):781-6.


Small interfering synthetic double-stranded RNA (siRNA) was applied to suppress the expression of the human cytotoxic- T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) gene transformed in transgenic rice cell cultures. The sequence of the 21-nucleotide siRNA was deliberately designed and synthesized with overhangs to inactivate the expression of hCTLA4Ig. The chemically synthesized siRNA duplex was combined with polyethyleneimine (PEI) at a mass ratio of 1:10 (0.33 microg siRNA:3.3 microg PEI) to produce complexes. The siRNA complexes (siRNA+PEI) were labeled with Cy3 in order to subsequently confirm the delivery by fluorescent microscopy. In addition, the cells were treated with sonoporation at 40 kHz and 419 W for 90 s to improve the delivery. The siRNA complexes alone inhibited the expression of hCTLA4Ig to 45% compared with control. The siRNA complexes delivered with sonoporation downregulated the production of hCTLA4Ig to 73%. Therefore, we concluded that the delivery of siRNA complexes into plant cells could be enhanced successfully by sonoporation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / genetics
  • Antigens, CD / metabolism
  • CTLA-4 Antigen
  • Cell Culture Techniques
  • Cell Survival
  • Gene Transfer Techniques*
  • Humans
  • Oryza / cytology
  • Oryza / genetics
  • Oryza / metabolism*
  • Plants, Genetically Modified / cytology
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism
  • RNA Interference*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism


  • Antigens, CD
  • CTLA-4 Antigen
  • CTLA4 protein, human
  • RNA, Small Interfering
  • Recombinant Proteins