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

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K15775
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 43060:System genome science-related
• Research Institution: University 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
• 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

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.

Free Research Field

バイオインフォマティクス

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。