Ultra efficient knockin system

• Project/Area Number: 16K07085
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: AIDA Tomomi 東京医科歯科大学, 難治疾患研究所, 准教授 (50540481)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: CRISPR / ノックイン / マウス / long ssDNA / PITCh / MMEJ / Exo1

Outline of Final Research Achievements

Precise and highly efficient integration of the exogenous gene into the target genomic locus (knockin) in vivo is the main challenge in the genome editing field. We have developed a series of new CRISPR technologies that enable highly efficient (up to 100%) knockin in mouse embryos.

Academic Significance and Societal Importance of the Research Achievements

ゲノム編集技術を用いて、あらゆる遺伝子配列を極めて高効率に動物個体に正確に導入可能にした。これらはヒト疾患に忠実な動物モデルの確立、高効率で精度の高い遺伝子治療法などの基盤として世界中で用いられている。

Research Products

• [Journal Article] Biased genome editing using the local accumulation of DSB repair molecules system. (2018)
  • Author(s): Nakade S, Mochida K, Kunii A, Nakamae K, Aida T, Tanaka K, Sakamoto N, Sakuma T, Yamamoto T
  • Journal Title: Nature Communications Volume: 9(1) Issue: 1 Pages: 3270-3270
  • DOI: 10.1038/s41467-018-05773-6
  • Peer Reviewed / Open Access
• [Journal Article] DEPDC5 deficiency contributes to resistance to leucine starvation via p62 accumulation in hepatocellular carcinoma (2018)
  • Author(s): Mizuno Y, Shimada S, Akiyama Y, Watanabe S, Aida T, Ogawa K, Ono H, Mitsunori Y, Ban D, Kudo A, Arii S, Yamaoka S, Tanabe M, Tanaka S
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 106-106
  • DOI: 10.1038/s41598-017-18323-9
  • Peer Reviewed / Open Access
• [Journal Article] Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins (2017)
  • Author(s): Quadros RM, Miura H, Harms DW, Akatsuka H, Sato T, Aida T, Redder R, Richardson GP, Inagaki Y, Sakai D, Buckley SM, Seshacharyulu P, Batra SK, Behlke MA, Zeiner SA, Jacobi AM, Izu Y, Thoreson WB, Urness LD, Mansour SL, Ohtsuka M, Gurumurthy CB
  • Journal Title: Genome Biology Volume: 18 Issue: 1 Pages: 92-92
  • DOI: 10.1186/s13059-017-1220-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Impaired degradation of medullary WNK4 in the kidneys of KLHL2 knockout mice. (2017)
  • Author(s): Kasagi Y, Takahashi D, Aida T, Nishida H, Nomura N, Zeniya M, Mori T, Sasaki E, Ando F, Rai T, Uchida S, Sohara E.
  • Journal Title: Biochem Biophys Res Commun. Volume: 487 Issue: 2 Pages: 368-374
  • DOI: 10.1016/j.bbrc.2017.04.068
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ. (2016)
  • Author(s): Aida T, Nakade S, Sakuma T, Izu Y, Oishi A, Mochida K, Ishikubo H, Usami T, Aizawa H, Yamamoto T, Tanaka K.
  • Journal Title: BMC Genomics Volume: 17 Issue: 1 Pages: 979-979
  • DOI: 10.1186/s12864-016-3331-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 受精卵ノックインの分子機構 (2018)
  • Author(s): 相田知海
  • Organizer: 日本ゲノム編集学会
  • Invited
• [Presentation] Making knock-in mice easy with Cloning-free CRISPR (2017)
  • Author(s): Tomomi Aida
  • Organizer: JSGE Satellite Symposium
  • Int'l Joint Research / Invited
• [Presentation] Opportunities and challenges in animal model innovation in CRISPR-era (2017)
  • Author(s): 相田知海
  • Organizer: 日本神経化学会
  • Invited