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     About RNAi Fly

Introduction
  A comprehensive RNAi-mutant fly bank has been constructed to provide tools for better understanding of gene functions, genetic networks, screening for new genes and so on. Project started on 2001, supported by the grant of National bio-Resource Project (NBRP) in Japan, and we started to release 1,000 lines to the community on 2005. We have been releasing stocks step-by-step and now there are about 11,000 lines on our on line ordering system. It would be our honor if these RNAi stocks can contribute to the basic science study.

  Using UAS-GAL4 system, the RNAi flies produce double stranded RNA in vivo ,ginducible RNAih. Even though this technique is not fully understood, its usefulness has been proven in many aspects of fly genetics. Users are referred to gDirections for Useh for technical advice. It is our hope that these RNAi flies will contribute to the development of basic science research.   

  We especially appreciate the following supports from collaborators, the Genetic Informatics Lab at NIG for web site construction, DNA Data Analysis lab at NIG for joint sequence program production, K. Saigo's lab at Univ. of Tokyo for primer selection and in silico sequence production, S. Morishita's lab at Univ. of Tokyo for off-target information production, and Mitsubishi Kagaku Institute of Life Sciences (MITILIS) for deposition of fly stocks. The NIG-Fly stock center is supported by NBRP grant from MEXT in Japan.

  We would like to request to add an acknowledgement when publishing your research work with these stocks. We suggest the following statement to be used: "The fly stock was obtained from National Institute of Genetics Fly Stock Center." Please send hard copies or reprints of all publications resulting from the use of the stocks and/or email us PDF files at flyadmin@lab.nig.ac.jp.

(April 2008)
Ryu Ueda
National Institute of Genetics (NIG), Genetic Strains Research Center


 Method
  • Primer design
    •   For PCR cloning, optimal primers were designed to cover a fragment at least 350 bp to 500 bp long near the 5f end of each ORF. Since some of the cloned fragment has homology against other genes, please see gDirections for useh for possibility of cross-reactions.
  • Vector construction and cloning strategies
    •   pUAST-R57: The MCS of the pUAST vector was modified for cloning inverted repeat (IR) cDNA fragment. Ret oncogene fragment, exon 5 to 7 including intron, was placed between IR fragments to enhance RNAi deficiency. For cloning strategies, please see Fig.1.
  • Injection and line establishment
    •   IR vector DNA 50 ng and helper (phs-pai) DNA 50 ng were transformed into 30 fly eggs. About half of them developed to adult flies. We used four transformed adults to establish IR fly lines by traditional genetic methods. For mating scheme, please see Fig.2.


Directions for use
  • white+ is used as a marker but some lines are contaminated with white-eyed flies. In most case, white-eyed flies are originated from a failure of balancing caused by multiple insertions. However, there are some cases, which we cannot explain. We are trying to cure these lines, anyway please check their eye color before using.
     
  • CAUTION!! There might be cross-reactions between IR fragment and off-target genes. Please refer goff-target informationh in each stock details to check cross-reaction possibilities.
     
  • Many lines appear lethal when RNAi is induced by Act5C-GAL4 at 28 degrees. This may be a detrimental effect of strong ubiquitous expression of RNAi. To interpret the effects of RNAi, you might a) use tissue specific drivers, b) compare the RNAi effect with the phenotypes of mutations in the gene, and c) consider using other RNAi flies such as VDRC (Vienna Drosophila RNAi Center: http://stockcenter.vdrc.at/control/main).
     
  • RNAi causes a hypomorophic effect, i.e. gene activity does NOT become null. Increasing the copy number of IR vectors might produce a stronger phenotype.
     
  • We CANNOT guarantee that our stocks are free of mites. DO NOT mix with your clean stocks before inspection.
     


Contributed Members
  • National Institute of Genetics (NIG)
    • Invertebrate Genetics Laboratory
      TAKAHASHI Kuniaki, TANIGUCHI Misako, SADO Yukiko, FUJITANI Kazuko
      KATO Naoko, HATANAKA Hiromi, NAKAGUTCHI Kaeko
      KAGOSHIMA Yuki, MASUNARI Miki, KOBARI Miyuki, KUMAGAI Niji, MOCHIZUKI Kaori, OCHIAI Michiko, HOSOKAWA Mayumi, SUZUKI Satomi, TASHIRO Yumiko, NAKANO Mariko
      OHYABU Yoko, SUZUKI Keiko, MASE Reiko, SAKODA Misako, SUDA Ritsuko, MARUYAMA Noriko
  • National Institute of Genetics (NIG)
    • Genetic Informatics Laboratory (http://www.shigen.nig.ac.jp/)
      YAMAZAKI Yukiko, YAMAKAWA Takehiro, YAHAGI Wataru, OOGUSHI Kazuhioro, WATANABE Koji, WATANABE Toru, KIMURA Gaku
  • National Institute of Genetics (NIG)
  • Mitsubishi Kagaku Institute of Life Sciences (MITILS)
    • (http://www.mitils.co.jp/)
      AWANO Wakae, OHTSU Kanae, YAMAMOTO Michiko, JUNI Aya, HASHIMOTO Naoko, YAMASHITA Atsushi
      SHIMAMURA Rieko, TATSUMI Ryoko, DOI Asako, MORIKAWA Yumiko, SATO Shobu, ISHIKAWA Shin-ichi
      AMANO Chiyoko, ITO Kiyomi, YODA Naomi
      GOTO Satoshi, ABE Masato
  • The University of Tokyo
    • Saigo Lab
      SAIGO Kaoru, TEI-UI Kumiko
      ZENNO Syuhei, KOJIMA Tetsuya, TAKAHASHI Fumitaka
      NAITO Yuki
  • The University of Tokyo
  • DYNACOM


Contact US
Administrator
Invertebrate Genetics Laboratory,
Genetic Strains Research Center,
National Institute of Genetics (NIG)
Mishima, Shizuoka, 411-8540, Japan
Phone: +81-55-981-6824
Fax: +81-55-981-6825
flyadmin@lab.nig.ac.jp

SHIGEN - SHared Information of GENetic resources - National Institute of Genetics (NIG)
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All rights reserved.
The NIG-Fly is a part of the National BioResource Project which started in 2002.