Establishment of stable hFis1 knockdown cells with an siRNA expression vector

被引：7

作者：

Arai, R

Ito, K

Wakiyama, M

Matsumoto, E

Sakamoto, A

Etou, Y

Otsuki, M

Inoue, M

Hayashizaki, Y

Miyagishi, M

Taira, K

Shirouzu, M

Yokoyama, S

机构：

[1] RIKEN, Genom Sci Ctr, Prot Res Grp, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan

[2] RIKEN, Genom Sci Ctr, Genome Explorat Res Grp, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan

[3] RIKEN, Harima Inst SPring 8, Structurome Res Grp, Sayo, Hyogo 6795148, Japan

[4] RIKEN, Harima Inst SPring 8, Struct & Mol Biol Lab, Sayo, Hyogo 6795148, Japan

[5] Univ Tokyo, Grad Sch Engn, Dept Chem & Biotechnol, Bunkyo Ku, Tokyo 1138656, Japan

[6] AIST, Funct Res Ctr, Tsukuba, Ibaraki 3058562, Japan

[7] Univ Tokyo, Grad Sch Sci, Dept Biophys & Biochem, Bunkyo Ku, Tokyo 1130033, Japan

来源：

JOURNAL OF BIOCHEMISTRY | 2004年 / 136卷 / 04期

关键词：

Fis1; knockdown; mitochondrial fission; RNAi; siRNA;

D O I：

10.1093/jb/mvh139

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Yeast Fis1p participates in mitochondrial fission, together with Dnm1p and Mdv1p. Recently, human Fis1 (hFis1) was reported to be involved in mitochondrial fission, together with Drp1. We established stable transformants with an hFis1 siRNA expression vector. In the stable hFis1 knockdown cells, hFis1 expression was suppressed to approximately 10%, and mitochondrial fission, induced by cisplatin treatment, was delayed. In addition, mouse Fis1 (mFis1) expression promoted mitochondrial fission and cell death in the hFis1 knockdown cells, suggesting that mFis1 complements the function of hFis1. These hFis1 siRNA expression vectors may be useful for studying the molecular function of mammalian Fis1.

引用

页码：421 / 425

页数：5

共 22 条

[1] A system for stable expression of short interfering RNAs in mammalian cells [J].

Brummelkamp, TR ;

Bernards, R ;

Agami, R .

SCIENCE, 2002, 296 (5567) :550-553

[2]

FRANCIS B, 2001, SEX ED, V1, P1

[3] hFis1, a novel component of the mammalian mitochondrial fission machinery [J].

James, DI ;

Parone, PA ;

Mattenberger, Y ;

Martinou, JC .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (38) :36373-36379

[4] Functional annotation of a full-length mouse cDNA collection [J].

Kawai, J ;

Shinagawa, A ;

Shibata, K ;

Yoshino, M ;

Itoh, M ;

Ishii, Y ;

Arakawa, T ;

Hara, A ;

Fukunishi, Y ;

Konno, H ;

Adachi, J ;

Fukuda, S ;

Aizawa, K ;

Izawa, M ;

Nishi, K ;

Kiyosawa, H ;

Kondo, S ;

Yamanaka, I ;

Saito, T ;

Okazaki, Y ;

Gojobori, T ;

Bono, H ;

Kasukawa, T ;

Saito, R ;

Kadota, K ;

Matsuda, H ;

Ashburner, M ;

Batalov, S ;

Casavant, T ;

Fleischmann, W ;

Gaasterland, T ;

Gissi, C ;

King, B ;

Kochiwa, H ;

Kuehl, P ;

Lewis, S ;

Matsuo, Y ;

Nikaido, I ;

Pesole, G ;

Quackenbush, J ;

Schriml, LM ;

Staubli, F ;

Suzuki, R ;

Tomita, M ;

Wagner, L ;

Washio, T ;

Sakai, K ;

Okido, T ;

Furuno, M ;

Aono, H .

NATURE, 2001, 409 (6821) :685-690

[5] Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells [J].

Lee, NS ;

Dohjima, T ;

Bauer, G ;

Li, HT ;

Li, MJ ;

Ehsani, A ;

Salvaterra, P ;

Rossi, J .

NATURE BIOTECHNOLOGY, 2002, 20 (05) :500-505

[6] Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method [J].

Livak, KJ ;

Schmittgen, TD .

METHODS, 2001, 25 (04) :402-408

[7] U6 promoter-driven siRNAs with four uridine 3′ overhangs efficiently suppress targeted gene expression in mammalian cells [J].

Miyagishi, M ;

Taira, K .

NATURE BIOTECHNOLOGY, 2002, 20 (05) :497-500

[8] Strategies for generation of an siRNA expression library directed against the human genome [J].

Miyagishi, M ;

Taira, K .

OLIGONUCLEOTIDES, 2003, 13 (05) :325-333

[9] Dnm1p GTPase-mediated mitochondrial fission is a multi-step process requiring the novel integral membrane component Fis1p [J].

Mozdy, AD ;

McCaffery, JM ;

Shaw, JM .

JOURNAL OF CELL BIOLOGY, 2000, 151 (02) :367-379

[10] Analysis of the mouse transcriptome based on functional annotation of 60,770 full-length cDNAs [J].

Okazaki, Y ;

Furuno, M ;

Kasukawa, T ;

Adachi, J ;

Bono, H ;

Kondo, S ;

Nikaido, I ;

Osato, N ;

Saito, R ;

Suzuki, H ;

Yamanaka, I ;

Kiyosawa, H ;

Yagi, K ;

Tomaru, Y ;

Hasegawa, Y ;

Nogami, A ;

Schönbach, C ;

Gojobori, T ;

Baldarelli, R ;

Hill, DP ;

Bult, C ;

Hume, DA ;

Quackenbush, J ;

Schriml, LM ;

Kanapin, A ;

Matsuda, H ;

Batalov, S ;

Beisel, KW ;

Blake, JA ;

Bradt, D ;

Brusic, V ;

Chothia, C ;

Corbani, LE ;

Cousins, S ;

Dalla, E ;

Dragani, TA ;

Fletcher, CF ;

Forrest, A ;

Frazer, KS ;

Gaasterland, T ;

Gariboldi, M ;

Gissi, C ;

Godzik, A ;

Gough, J ;

Grimmond, S ;

Gustincich, S ;

Hirokawa, N ;

Jackson, IJ ;

Jarvis, ED ;

Kanai, A .

NATURE, 2002, 420 (6915) :563-573

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