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Molecular, Biochemical, and Genetic Techniques

開催日 2014/9/12
時間 11:00 - 12:00
会場 Poster / Exhibition(Event Hall B)

高速・高効率in vivoゲノム編集によるノックインマウス作出
Rapid and highly efficient in vivo genome editing for knockin mouse: from human rare variants to gene cassettes

  • P2-381
  • 今橋 里沙 / Risa Imahashi:1 相田 知海 / Tomomi Aida:1 張 景ほう / Keiho Chiyo:1 佐久間 哲史 / Tetsushi Sakuma:5 宇佐美 貴子 / Takako Usami:2 石久保 春美 / Harumi Ishikubo:1 Pawel Obrocki / Obrocki Pawel:1 山本 卓 / Takashi Yamamoto:5 田中 光一 / Kohichi Tanaka:1,3,4 
  • 1:東医歯大難治疾患研分子神経科学分野 / Mol Neurosci, Med Res Inst, Tokyo Med Dent Univ, Tokyo 2:遺伝子組換えマウス実験室 / Lab of Reco Ani, Med Res Inst 3:東医歯大脳統合機能研セ / Cent Brain Integ Res, Tokyo Med Dent Univ, Tokyo 4:科学技術振興機構 CREST / JST, CREST, Saitama, Japan 5:広島大院理数理分子生命理学 / Dept of Math and Life Sci, Grad Sch of Sci, Hiroshima Univ , Hiroshima 

Gene targeting in mouse has dramatically improved our understanding of the functions of genes in vivo in health and diseases. However, it is a time-consuming, laborious, and expensive process. The recent groundbreaking development of genome editing technologies has enabled direct manipulation of the genome in mouse zygotes (in vivo genome editing), thereby providing new avenues for simple, convenient, highly efficient and ultra-rapid production of knockout or knockin mice without ES cells. We focus on functional significance of multiple human rare variants in glutamate transporter EAAT1 found in glaucoma patients. To achieve this in vivo, we have generated several knockin mouse lines carrying precisely modified these rare variants with highly active Platinum TALENs and oligo DNA donors. We have also generated knockin mice carrying a 6.5kb gene cassette containing EGFP reporter with CRISPR/Cas and targeting vector for fine tracing of specific-cell lineage. Because the efficiency of our method is quite high, several germline-competent knockin founders can be obtained within a month by single microinjection. Taken together, in vivo genome editing technology revolutionizes mouse gene targeting and provides exciting opportunities for the functional genomic and neurodevelopmental researches in vivo.

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