title: Enhanced delivery of siRNA complexes by sonoporation in transgenic rice cell suspension cultures.
abstract: 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.
authors.affiliation: Department of Biological Engineering, Inha University, Incheon 402-751, Korea.
authors.forename: Su-Hwan
authors.initials: SH
authors.lastname: Cheon
authors.forename: Kyoung-Hoon
authors.initials: KH
authors.lastname: Lee
authors.forename: Jun-Young
authors.initials: JY
authors.lastname: Kwon
authors.forename: Sung-Hun
authors.initials: SH
authors.lastname: Choi
authors.forename: Mi-Na
authors.initials: MN
authors.lastname: Song
authors.forename: Dong-Il
authors.initials: DI
authors.lastname: Kim
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D060908:CTLA-4 Antigen; D018929:Cell Culture Techniques; D002470:Cell Survival; D018014:Gene Transfer Techniques; D006801:Humans; D012275:Oryza / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D034741:RNA, Small Interfering; D011994:Recombinant Proteins