Development of a novel genome editing-mediated gene knock-in system

2016 Fiscal Year Final Research Report

• Project/Area Number: 26290070
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: System genome science
• Research Institution: Hiroshima University
• Principal Investigator: Yamamoto Takashi 広島大学, 理学(系)研究科(研究院), 教授 (90244102)
• Co-Investigator(Kenkyū-buntansha): 谷口 俊介 筑波大学, 生命環境科学研究科(系), 准教授 (00505331)
• Co-Investigator(Renkei-kenkyūsha): AIDA TOMOMI 東京医科歯科大学, 難治疾患研究所, 准教授 (50540481) ; SAKAMOTO NAOAKI 広島大学, 大学院理学研究科, 准教授 (00332338) ; SUZUKI KENICHI 広島大学, 大学院理学研究科, 特任准教授 (90363043)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: バイオテクノロジー / ゲノム編集

Outline of Final Research Achievements

In this study, I established microhomology-mediated end-joining (MMEJ) dependent gene knock-in system that I termed precise integration into target chromosome (PITCh). I performed reporter gene knock-in experiments using PITCh system in cultured cells and animals such as frog and mice. As results, I observed efficient and precise gene knock-in by PITCh method compared with those achieved by HR-mediated gene knock-in. Additionally, I performed genetic screening of MMEJ enhancers and identified that PITCh efficiency upon Exonuclease I (ExoI) overexpression was increased to 2.5 fold compared with that upon mock overexpression. I also confirmed the effect of ExoI overexpression in endogenous gene locus in cultured cells and mouse zygote.

Free Research Field

ゲノム生物学