| Budget Amount *help |
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2016: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2015: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2014: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
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| 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.
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