Soaking RNAi in Bombyx mori BmN4-SID1 cells arrests cell cycle progression.
RNA interference (RNAi) is an evolutionarily conserved mechanism for sequence-specific gene silencing. Previously, the BmN4-SID1 cell expressing Caenorhabditis ele gans SID-1 was established, in which soaking RNAi could induce effective gene silencing. To establish its utility, 6 cell cycle progression related cDNAs, CDK1, MYC, MYB, RNRS, CDT1, and GEMININ, were isolated from the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), and their expressions were further silenced by soaking RNAi in the BmN4-SID1 cells. The cell cycle progression analysis using flow cytometer demonstrated that the small amount of double stranded RNA was enough to arrest cell cycle progression at the specific cell phases. These data suggest that RNAi in the BmN4-SID1 cells can be used as a powerful tool for loss-of-function analysis of B. mori genes.
1 Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Hakozaki 6-10-1, Fukuoka 812-8581.
Hiroaki
H
Mon
Zhiqing
Z
Li
Isao
I
Kobayashi
Shuichiro
S
Tomita
JaeMan
J
Lee
Hideki
H
Sezutsu
Toshiki
T
Tamura
Takahiro
T
Kusakabe
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000502:physiology; D012831:Bombyx / Q000166:cytology* / Q000235:genetics / Q000502:physiology*; D017173:Caenorhabditis elegans; D002453:Cell Cycle; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017423:Sequence Analysis, RNA
D018076:DNA, Complementary; D019476:Insect Proteins; D012330:RNA, Double-Stranded