Application of lipofection-mediated genome editing in mouse zygotes created by IVF via ultra-superovulation

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H04606
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Integrative animal science
• Research Institution: Kumamoto University
• Principal Investigator: Nakagata Naomi 熊本大学, 生命資源研究・支援センター, 教授 (30159058)
• Co-Investigator(Kenkyū-buntansha): 中川 佳子 熊本大学, 生命資源研究・支援センター, 特任助教 (30732739)
• Co-Investigator(Renkei-kenkyūsha): Yamamoto Takashi 広島大学, 大学院理学研究科, 教授 (90244102) ; Sakuma Tetsushi 広島大学, 大学院理学研究科, 講師 (90711143)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: マウス / ゲノム編集 / 受精卵 / 核酸導入

Outline of Final Research Achievements

Recently, genome editing in mouse zygotes has become convenient and scalable, in association with various technological developments and improvements such as novel nuclease tools, alternative delivery methods, and contemporary reproductive engineering techniques. We have so far demonstrated the applicability of ultra-superovulation, in vitro fertilization (IVF), and vitrification/warming of zygotes in microinjection-mediated mouse genome editing. Here, we present an example of an application coupling the following three technologies: 1) CRISPR-Cas9 system as the most convenient genome-editing technology, 2) lipofection as the most simple, economical and effortless delivery method, and 3) cryopreserved oocytes created by IVF via ultra-superovulation as the most animal welfare- and user-friendly strategy. We successfully created gene knockout mice carrying insertion/deletion mutations.

Free Research Field

生殖工学