Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1

Hiroaki Mon; Isao Kobayashi; Shinji Ohkubo; Shuichiro Tomita; Jae Man Lee; Hideki Sezutsu; Toshiki Tamura; Takahiro Kusakabe

doi:10.4161/rna.9.1.18084

Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1

Hiroaki Mon, Isao Kobayashi, Shinji Ohkubo, Shuichiro Tomita, Jae Man Lee, Hideki Sezutsu, Toshiki Tamura, Takahiro Kusakabe

Agricultural Bioresource Sciences

Research output: Contribution to journal › Article › peer-review

63 Citations (Scopus)

Abstract

RNA interference (RNAi) is a conserved mechanism that catalyzes sequence-specific gene silencing, and has been used for loss-of-function genetic screens in many organisms. Here, we demonstrated that the expression of Caenorhabditis elegans SID-1 (CeSID-1) could trigger effective gene silencing in the cultured silkworm cell line, BmN4 (BmN4-SID1). Soaking the BmN4-SID1 in dsRNA corresponding to endogenous target genes induced a significant decrease of the amount of mRNA or protein. A small amount of dsRNA was enough to silence the target gene in a few days. Overexpression of CeSID-1 did not affect the cell viability. Our results suggest that BmN4-SID1 can be used in many applications in silkworm cells and will become a valuable resource for gene analysis.

Original language	English
Pages (from-to)	40-46
Number of pages	7
Journal	RNA biology
Volume	9
Issue number	1
DOIs	https://doi.org/10.4161/rna.9.1.18084
Publication status	Published - Jan 2012

All Science Journal Classification (ASJC) codes

Molecular Biology
Cell Biology

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title = "Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1",

abstract = "RNA interference (RNAi) is a conserved mechanism that catalyzes sequence-specific gene silencing, and has been used for loss-of-function genetic screens in many organisms. Here, we demonstrated that the expression of Caenorhabditis elegans SID-1 (CeSID-1) could trigger effective gene silencing in the cultured silkworm cell line, BmN4 (BmN4-SID1). Soaking the BmN4-SID1 in dsRNA corresponding to endogenous target genes induced a significant decrease of the amount of mRNA or protein. A small amount of dsRNA was enough to silence the target gene in a few days. Overexpression of CeSID-1 did not affect the cell viability. Our results suggest that BmN4-SID1 can be used in many applications in silkworm cells and will become a valuable resource for gene analysis.",

author = "Hiroaki Mon and Isao Kobayashi and Shinji Ohkubo and Shuichiro Tomita and Lee, {Jae Man} and Hideki Sezutsu and Toshiki Tamura and Takahiro Kusakabe",

note = "Funding Information: This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) 5{\textquoteright}-GCGTAATACGACTCACTATAGGGGCCGCCAG- and by a grant from the Ministry of Agriculture, Forestry and TGTGCTGGAATTCTGCAGAT-3{\textquoteright} Fisheries of Japan (Integrated research project for plant, insect 5{\textquoteright}-GCGTAATACGACTCACTATAGGGTGCTCGA- and animal using genome technology INSECT-1201), and Do not disKAKEtNHIrno.ib22248003utande22248004. from the Japan ForRT-PCR Society for the Promotion of Science. We thank Dr. Aoki (Kyushu University Graduate School) for providing the BmN4 cell line.",

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doi = "10.4161/rna.9.1.18084",

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AU - Mon, Hiroaki

AU - Kobayashi, Isao

AU - Ohkubo, Shinji

AU - Tomita, Shuichiro

AU - Lee, Jae Man

AU - Sezutsu, Hideki

AU - Tamura, Toshiki

AU - Kusakabe, Takahiro

N1 - Funding Information: This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) 5’-GCGTAATACGACTCACTATAGGGGCCGCCAG- and by a grant from the Ministry of Agriculture, Forestry and TGTGCTGGAATTCTGCAGAT-3’ Fisheries of Japan (Integrated research project for plant, insect 5’-GCGTAATACGACTCACTATAGGGTGCTCGA- and animal using genome technology INSECT-1201), and Do not disKAKEtNHIrno.ib22248003utande22248004. from the Japan ForRT-PCR Society for the Promotion of Science. We thank Dr. Aoki (Kyushu University Graduate School) for providing the BmN4 cell line.

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Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1

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