title: [Evaluation of transfection effectiveness using fluorescein-labelled oligonucleotides and entraster-R siRNA transfection into Plasmodium falciparum].
abstract: The cultured Plasmodium falciparum parasites were synchronized twice by 5% sorbitol treatment twice (8-hour window), and then incubated at 37 degrees C for 16 h. Parasites were transfected with fluorescein-labelled oligonucleotides (group A) or fluorescein-labelled oligonucleotides+Entranster-R siRNA transfection reagent (group B). After 5 h a part of parasites was evaluated by fluorescence microscopy and flow cytometry. The rest of parasites were washed with RPMI 1640 medium, and then incubated with 500 microl new medium containing 2% fresh erythrocytes for another 12 h, and detected by flow cytometry. The fluorescein-labelled oligonucleotides were localized in erythrocytes in group B, but nearly no fluorescence was observed for group A. Flow cytometry analysis indicated that the transfection efficiency of group B [(47.40 +/- 3.39)%] was higher than that of group A [(0.60 +/- 0.27)%]. In the second cell cycle, the transfection efficiency in group B was (26.85 +/- 2.90)%, while that of group A was nearly zero. The results indicated that Entranster-R siRNA transfection reagent may increase the oligonucleotides transfection efficiency.
authors.forename: Hong-Chang
authors.initials: HC
authors.lastname: Zhou
authors.forename: Yu-Hui
authors.initials: YH
authors.lastname: Gao
authors.forename: Sheng-Wen
authors.initials: SW
authors.lastname: Shao
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
mesh_terms: D019793:Fluorescein; D009841:Oligonucleotides; D010963:Plasmodium falciparum / Q000235:genetics*; D034741:RNA, Small Interfering; D014162:Transfection / Q000379:methods*
chemical_list: D009841:Oligonucleotides; D034741:RNA, Small Interfering; D019793:Fluorescein