Development of mRNA ablation using completely new functional nucleic acid constructs

• Project/Area Number: 23501304
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Clinical oncology
• Research Institution: Niigata University
• Principal Investigator: NARITA Miwako 新潟大学, 医歯学系, 准教授 (30281009)
• Co-Investigator(Kenkyū-buntansha): NASHIMOTO Masayuki 新潟薬科大学, 応用生物科学部, 教授 (30228069)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: sgRNA / mRNA / apoptosis / tRNaseZL / WT1 / アポトーシス / 腫瘍細胞 / Bcl-2 / 白血病

Research Abstract

TRUE gene silencing is a new method to delete specific cellular RNAs by using tRNase ZL and small guide RNA (sgRNA) that cleaves a pre-tRNA-like or micro-pre-tRNA-like complex consisting of cellular target RNA. We investigated the possibility that heptamer-type sgRNA can be used as nucleic acid medicine to treat hematological malignancies. We investigated the effect of artificial sgRNAs on mRNAs which -targeting sgRNAs affect leukemic cells. We confirmed that sgRNA can be taken up by cells without any transfection reagents, and that the naked sgRNAs targeting several tumor-associated mRNA can reduce specific mRNA levels and the amounts of tumor-associated protein in several leukemic cell lines. These sgRNAs efficiently induced apoptosis in these cells.
These encouraging findings lead us to follow the possibility that heptamer-type sgRNAs targeting anti-apoptotic mRNAs can be used as RNA medicine against hematological malignancies.

Research Products

• [Journal Article] naked RNA heptamer targeting the human Bcl-2 mRNA induces apoptosis of HL60 leukemia cells (2013)
  • Author(s): Takahashi M, Elbarbary RA, Nakashima A, Abe M, Watanabe N, Narita M, Takahashi M, Tamura M, Yoshida T, Nashimoto M
  • Journal Title: Cancer Letters Volume: 328(2) Pages: 362-368
• [Journal Article] Induction of apoptosis of leukemic cells by TRUE gene silencing using small guide RNAs targeting the WT1 mRNA (2013)
  • Author(s): Watanabe N, Narita M, Saito A, Yamahira A, Taniguchi T, Furukawa T, Yoshida T, Miyazawa T, Nashimoto M, Takahashi M
  • Journal Title: Leuk Res Volume: 37(5) Pages: 580-585
• [Journal Article] Induction of apoptosis of leukemic cells by TRUE gene silencing using small guide RNAs targeting the WT1 mRNA. (2013)
  • Author(s): Watanabe N, Narita M, Yamahira A, Taniguchi T, Furukawa T, Yoshida T, Miyazawa T, Nashimoto M, Takahashi M.
  • Journal Title: Leuk Res. Volume: 37 Issue: 2 Pages: 580-5
  • DOI: 10.4081/rt.2011.e24
  • Peer Reviewed
• [Journal Article] A naked RNA heptamer targeting the human Bcl-2 mRNA induces apoptosis of HL60 leukemia cells (2013)
  • Author(s): Takahashi M, Elbarbary RA, Nakashima A, Abe M, Watanabe N, Narita M, Takahashi M, Tamura M, Yoshida T, Nashimoto M.
  • Journal Title: Cancer Lett. Volume: 328 Pages: 362-8
  • DOI: 10.1155/2010/623593
  • Peer Reviewed
• [Patent(Industrial Property Rights)] ヒト白血病細胞のアポトーシスを誘導するヘプタマー型スモールガイド核酸 (2012)
  • Inventor(s): 梨本正之、高橋益廣、成田美和子、吉田哲郎、宮澤達也
  • Industrial Property Rights Holder: 学校法人新潟科学技術学園新潟薬科大学、国立大学法人新潟大学
  • Industrial Property Rights Type: 特許
  • Acquisition Date: 2012-08-24
  • Overseas