Approach to allele specific and regulated gene silencing using the artificial miRNA expression system

• Project/Area Number: 21659084
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Human genetics
• Research Institution: Kyushu University
• Principal Investigator: FUKUMAKI Yasuyuki 九州大学, 生体防御医学研究所, 教授 (90128083)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,240,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2009: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: マイクQRNA / マイクQR / 遺伝子変異 / アレル特異的 / マイクロRNA / 発現抑制 / FAP / TTR / オフターゲット効果 / ミスマッチ

Research Abstract

The allele specific and regulated gene silencing is needed in various aspects of biological and medical fields. To establish this system, artificial miRNA driven by the poly II promoter was designed for discrimination between normal and mutated alleles in silencing effect. Although the allele specific silencing was obtained using artificial miRNA, there was no increased silencing effect expected to be provided by co-expression of another miRNA targeting the region shared by two genotypes. Cre dependent silencing of Smad2 and Smad3 as a model was obtained in ES cells using the construct in which expression of two miRNA species is blocked by the Neo^r gene flanked by two loxP sites.

Research Products

• [Journal Article] Partial SPAST and DPY30 deletions in a Japanese spastic paraplegia type 4 family (2011)
  • Author(s): Miura, S., Shibata, H., Kida, H., Noda, K., Toyama, T., Iwasaki, N., Iwaki, A., Ayabe, M., Aizawa, H., Taniwaki, T. and Fukumaki, Y.
  • Journal Title: Neurogenetics Volume: 12 Pages: 25-31
  • Peer Reviewed
• [Journal Article] Multiple system degeneration with basophilic inclusions in Japanese ALS patients with FUS mutation (2010)
  • Author(s): Tateishi, T., Hokonohara, T., Yamasaki, R., Miura, S., Kikuchi, H., Iwaki, A., Tashiro, H., Furuya, H., Nagara, Y., Ohyagi, Y., Nukina, N., Iwaki, T., Fukumaki, Y. and Kira, JI.
  • Journal Title: Ac to Neuropathol Volume: 119 Pages: 355-364
  • Peer Reviewed
• [Presentation] Linkageassisted exome sequencing to identify the responsible variant for a novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a single Japanese family (2012)
  • Author(s): Shibata, H., Miura, S., Kida, H., Neda, K., Kaku, Y., Iwaki, A., Ayabe, M., Taniwaki, T. and Fukumaki, Y.
  • Organizer: Human Genome Meeting 2012
  • Place of Presentation: Sydney, Australia
• [Presentation] Linkage-assisted exome sequencing to identify the responsible variant for a novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a single Japanese family (2012)
  • Author(s): Shibata, H., et al
  • Organizer: Human Genome Meeting 2012
  • Place of Presentation: Sydney, Australia
• [Presentation] Exome sequencing approach to identify the responsible variant for a novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a Japanese descent (2011)
  • Author(s): Shibata, H., Miura, S., Kida, H., Neda, K., Kaku, Y., Iwaki, A., Ayabe, M., Taniwaki, T. and Fukumaki, Y.
  • Organizer: The 12th International Congress of Human Genetics/The 61st Annual Meeting of The American Society of Human Genetics
  • Place of Presentation: Montreal, Canada
• [Presentation] A novel type of hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities found in a Japanese descent (2010)
  • Author(s): Miura, S., Shibata, H., Neda, K., Y. Kaku, Y. Kida, H., Iwaki, A., Ayabe, M., Aizawa, H., Taniwaki, T. and Fukumaki, Y.
  • Organizer: 135th Annual Meeting of American Neurological Association
  • Place of Presentation: San Francisco, CA, USA
• [Presentation] Haplotype analysis of fukutin(FICTN) gene in two very mild Fukuyama type congenital muscular dystrophy(FCMD) (2010)
  • Author(s): Furuya, H., Arahata, H., Fuji i, N., Miura, S., Shibata, H. and Fukumaki, Y.
  • Organizer: 135th Annual Meeting of American Neurological Association
  • Place of Presentation: San Francisco, CA, USA
• [Presentation] Partial SPAST and DPYso deletions in autosomal dominant hereditary spastic paraplegia (2010)
  • Author(s): Miura, S., Shibata, H., Kida, K. Neda, A. Iwaki, A., M. Ayabe, M., H. Aizawa, H., T. Taniwaki, T. and Fukumaki, Y. X
  • Organizer: 60th Annual Meeting of The American Society of Human Genetics
  • Place of Presentation: Washington, DC, USA
• [Presentation] Stoichiometric Analysis of Short Hairpin RNAs in Cultured Cells (2009)
  • Author(s): Shibata, A, et al.
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-12
• [Presentation] Stoichiometric analysis of short hairpin RNAs in cultured cells (2009)
  • Author(s): Shibata, A., Kaneko, S., Teramoto, N. and Fukumaki, Y.
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
• [Remarks] ホームページ
  • URL: http://www.gen.kyushu-u.ac.jp/Tbyouin/
• [Remarks]
  • URL: http://www.gen.kyushu-u.ac.jp/~byouin/
• [Remarks]
  • URL: http://www.gen.kyushu-u.ac.jp/~byouin/