Efficient gene silencing and cell differentiation using siRNA in mouse and monkey ES cells.
Small interfering RNA (siRNA) has been widely used for suppressing gene expression in various organisms. Here, we describe efficient methods to suppress target genes (EGFP or Oct4) using siRNA in mouse and monkey ES cells, and differentiation. In mouse ES cells, FACS analysis revealed that EGFP expression was suppressed in 97% of transfected cells at 48 h after transfection. In addition, cells expressed Hand1 and Cdx2, which are the marker genes of trophoblast lineage by the transient suppression of Oct4. In the case of monkey ES cells, highly efficient suppression was achieved in 98% of cells at 96 h post-transfection using the Sendai virus (hemagglutinating virus of Japan, HVJ) envelope as a carrier of siRNA. These efficient transfection methods using synthetic siRNA should contribute to evaluate specific gene function in ES cells and can be used to differentiate ES cells into desired cell lineages.
Takada
Tatsuyuki
T
Research Center for Animal Life Science, Shiga University of Medical Science, Ohtsu, Shiga 520-2192, Japan. ttakada@belle.shiga-med.ac.jp
Nemoto
Ken-ichi
K
Yamashita
Akihiro
A
Kato
Masaya
M
Kondo
Yasushi
Y
Torii
Ryuzo
R
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002478:Cells, Cultured; D017931:DNA Primers; D005434:Flow Cytometry; D020868:Gene Silencing; D049452:Green Fluorescent Proteins / Q000235:genetics; D000882:Haplorhini; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics*
D017931:DNA Primers; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins