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Development of a universal genome-editing genotyping method using a deep generation model

Research Project

Project/Area Number 20K15775
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 43060:System genome science-related
Research InstitutionUniversity of Tsukuba

Principal Investigator

Kuno Akihiro  筑波大学, 医学医療系, 助教 (60830122)

Project Period (FY) 2020-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2020: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywordsバイオインフォマティクス / ゲノム編集 / 実験動物学 / CRISPR-Cas / 次世代シーケンサー / 深層生成モデル
Outline of Research at the Start

CRISPR-Casによるゲノム編集技術の多方面における爆発的な普及の一方で、作出されたゲノム編集生物が目的の変異を持つ個体なのか、その品質を保証するために行う遺伝型解析手法では検出が困難な遺伝型が存在することが大きな問題となっている。
そこで本研究では多様なゲノム編集デザインに対して汎用的に用いることができる新規遺伝型解析手法の樹立を目的とし、ゲノム編集生物の質の保証という研究の根幹に貢献する。

Outline of Final Research Achievements

Although CRISPR-Cas revolutionized the efficiency of genome-editing mice's production, there has been no method to accurately and rapidly detect "unintended mutations" caused by genome editing. In this study, we developed Determine Allele mutations and Judge Intended genotypes by Nanopore sequencer (DAJIN), which can detect and classify "unintended mutations" and "intended mutations" caused by genome editing. DAJIN has demonstrated that it is capable of genotyping multiple types of genome editing, including point mutations, exon deletions, and induction of flox mutations, with greater accuracy than conventional methods.

Academic Significance and Societal Importance of the Research Achievements

意図した遺伝子改変マウス系統を実験に使用するためには、ゲノム編集を施された受精卵から発生した数十匹のマウスから「意図した改変遺伝子」を持つマウスを遺伝子検査で特定する必要がある。本研究で開発した遺伝子検査法(DAJIN)では、約100匹のマウスのそれぞれでどのような遺伝子改変が生じたのか、網羅的かつ正確に、簡便に特定できる。この手法はゲノム編集実験にかかる期間の短縮やその正確性の向上に寄与すると期待される。

Report

(3 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • Research Products

    (3 results)

All 2022 2021 2020

All Journal Article (2 results) (of which Peer Reviewed: 1 results,  Open Access: 2 results) Presentation (1 results)

  • [Journal Article] DAJIN enables multiplex genotyping to simultaneously validate intended and unintended target genome editing outcomes2022

    • Author(s)
      Kuno Akihiro, Ikeda Yoshihisa, Ayabe Shinya, et. al.
    • Journal Title

      PLOS Biology

      Volume: 20 Issue: 1 Pages: e3001507-e3001507

    • DOI

      10.1371/journal.pbio.3001507

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] DAJIN-assisted multiplex genotyping to validate the outcomes of CRISPR-Cas-based genome editing2020

    • Author(s)
      Kuno, A., Ikeda, Y., Ayabe, S., Kato, K., Sakamoto, K., Suzuki, S., ... & Mizuno, S.
    • Journal Title

      bioRxiv

      Volume: -

    • DOI

      10.1101/2020.12.14.422641

    • Related Report
      2020 Research-status Report
    • Open Access
  • [Presentation] DAJIN validates the multiallelic genome-editing outcomes using machine learning-assisted long-read sequencing2021

    • Author(s)
      久野 朗広
    • Organizer
      第10回生命医薬情報学連合大会(IIBMP2021)
    • Related Report
      2021 Annual Research Report

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Published: 2020-04-28   Modified: 2023-01-30  

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