title: Sendai F/HN viroplexes for efficient transfection of leukemic T cells.
abstract: OBJECTIVE
  Most chemical transfection reagents are ineffective for the transfection of cells in suspension, such as leukemic cell and stem cell lineages. We developed two different types of viroplexes, cationic Sendai F/HN viroplexes (CSVs) and protamine sulfate-condensed cationic Sendai F/HN viroplexes (PCSVs) for the efficient transfection of T-leukemic cells.
  
  
  METHODS
  The viroplex systems were prepared by reconstitution of fusogenic Sendai F/HN proteins in DMKE (O,O'-dimyristyl-N-lysyl glutamate) cationic liposomes. The viroplexes were further optimized for plasmid DNA and siRNA delivery to suspension cells. The particle size and surface charge of the viroplexes were analyzed with a ζ-sizer. Transfection of plasmid DNA (pDNA) and small interfering RNA (siRNA) by CSVs or PCSV was evaluated by measurement of transgene expression, confocal microscopy, FACS, and RT-PCR.
  
  
  RESULTS
  The optimized CSVs and PCSVs exhibited enhanced gene and siRNA delivery in the tested suspension cell lines (Jurkat cells and CEM cells), compared with conventional cationic liposomes. In the case of pDNA transfection, the CSVs and PCSVs show at least 10-fold and 100-fold higher transgene expression compared with DMKE lipoplexes (or lipofectamine 2000), respectively. The CSVs showed more effective siRNA delivery to the suspension cells than cationic liposomes, as assessed by confocal microscopy, FACS, and RT-PCR. The effective transfection by the CSVs and PCSVs is presumably due to fusogenic activity of F/HN proteins resulting in facilitated internalization of pDNA and siRNA.
  
  
  CONCLUSIONS
  This study suggests that Sendai F/HN viroplexes can be widely applicable for the transfection of pDNA and siRNA to suspension cell lines.
authors.affiliation: Department of Biomedical Laboratory Science, Yonsei University, 1 Yeonsedae-gil, Wonju 220-710, Korea.
authors.forename: Jung Seok
authors.initials: JS
authors.lastname: Kim
authors.forename: Yeon Kyung
authors.initials: YK
authors.lastname: Lee
authors.forename: Hwa Yeon
authors.initials: HY
authors.lastname: Jeong
authors.forename: Seong Jae
authors.initials: SJ
authors.lastname: Kang
authors.forename: Min Woo
authors.initials: MW
authors.lastname: Kim
authors.forename: Seung Hyun
authors.initials: SH
authors.lastname: Ryu
authors.forename: Hong Sung
authors.initials: HS
authors.lastname: Kim
authors.forename: Keun Sik
authors.initials: KS
authors.lastname: Kim
authors.forename: Dong-Eun
authors.initials: DE
authors.lastname: Kim
authors.forename: Yong Serk
authors.initials: YS
authors.lastname: Park
mesh_terms: D045744:Cell Line, Tumor; D015707:HN Protein / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells; D034741:RNA, Small Interfering; D029082:Sendai virus / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014760:Viral Fusion Proteins / Q000235:genetics; D022701:Virosomes
chemical_list: D015707:HN Protein; D034741:RNA, Small Interfering; D014760:Viral Fusion Proteins; D022701:Virosomes
keywords: Sendai F/HN protein; cationic Sendai viroplex; gene transfection; suspension cell
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