Mechanisms of DNA repair involved in the genome editing efficiencies in mouse zygotes

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K21654
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Integrative animal science; System genome science
• Research Institution: National Center for Child Health and Development
• Principal Investigator: Hara Satoshi 国立研究開発法人国立成育医療研究センター, システム発生・再生医学研究部, 研究員 (80739582)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: ゲノム編集

Outline of Final Research Achievements

Although CRISPR/Cas9-mediated genome editing technology is remarkably progressed, the insertion efficiencies of an exogenous sequence using this technology via the homology directed repair (HDR) are still low. In this study, to assess the relationships between the timing of HDR and cell cycle of zygotes, we microinjected sgRNA/Cas9 with oligomer containing exogenous sequences into mouse zygotes at the different pronucleus stages (PN1-2, PN2-3, PN3-4, PN4-5). The results showed that the highest insertion efficiencies were observed in the zygotes at the PN4-5 stage, whereas the lowest efficiencies in PN1-2 stage zygotes. From these results, HDR in mouse zygotes is also activated by the cell cycle-dependent manner.

Free Research Field

発生工学