Soaking RNAi inBombyx moriBmN4-SID1 Cells Arrests Cell Cycle Progression
Author(s) -
Hiroaki Mon,
Zhiqing Li,
Isao Kobayashi,
Shuichiro Tomita,
JaeMan Lee,
Hideki Sezutsu,
Toshiki Tamura,
Takahiro Kusakabe
Publication year - 2013
Publication title -
journal of insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.551
H-Index - 49
ISSN - 1536-2442
DOI - 10.1673/031.013.15501
Subject(s) - manduca sexta , 20 hydroxyecdysone , biology , ecdysone , nad+ kinase , sphingidae , midgut , manduca , ecdysone receptor , biochemistry , bombyx mori , intracellular , mitochondrion , microbiology and biotechnology , enzyme , botany , gene , insect , nuclear receptor , larva , transcription factor
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.
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