基本情報
- 所属
- 自治医科大学 先端医療技術開発センター 再生・細胞医薬研究ラボラトリー 講師
- 学位
- 農学(信州大学)
- J-GLOBAL ID
- 201601004312261838
- researchmap会員ID
- B000259287
経歴
4-
2020年4月 - 現在
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2016年3月 - 2020年3月
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2015年4月 - 2016年2月
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2013年4月 - 2015年3月
学歴
3-
2012年4月 - 2014年9月
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2010年4月 - 2012年3月
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2006年4月 - 2010年3月
受賞
1論文
30-
Molecular therapy. Methods & clinical development 20 451-462 2021年3月12日We conducted two lines of genome-editing experiments of mouse hematopoietic stem cells (HSCs) with the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9). First, to evaluate the genome-editing efficiency in mouse bona fide HSCs, we knocked out integrin alpha 2b (Itga2b) with Cas9 ribonucleoprotein (Cas9/RNP) and performed serial transplantation in mice. The knockout efficiency was estimated at approximately 15%. Second, giving an example of X-linked severe combined immunodeficiency (X-SCID) as a target genetic disease, we showed a proof-of-concept of universal gene correction, allowing rescue of most of X-SCID mutations, in a completely non-viral setting. We inserted partial cDNA of interleukin-2 receptor gamma chain (Il2rg) into intron 1 of Il2rg via non-homologous end-joining (NHEJ) with Cas9/RNP and a homology-independent targeted integration (HITI)-based construct. Repaired HSCs reconstituted T lymphocytes and thymuses in SCID mice. Our results show that a non-viral genome-editing of HSCs with CRISPR/Cas9 will help cure genetic diseases.
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Methods in molecular biology (Clifton, N.J.) 2320 247-259 2021年A knock-in can generate fluorescent or Cre-reporter under the control of an endogenous promoter. It also generates knock-out or tagged-protein with fluorescent protein and short tags for tracking and purification. Recent advances in genome editing with clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9) significantly increased the efficiencies of making knock-in cells. Here we describe the detailed protocols of generating knock-in mouse and human pluripotent stem cells (PSCs) by electroporation and lipofection, respectively.
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Experimental hematology 2021年1月1日We report that a sheep fetal liver provides a microenvironment for generating hematopoietic cells with long-term engrafting capacity and multilineage differentiation potential from human induced pluripotent stem cell (iPSC)-derived hemogenic endothelial cells (HEs). Despite the promise of iPSCs for making any cell types, generating hematopoietic stem and progenitor cells (HSPCs) is still a challenge. We hypothesized that the hematopoietic microenvironment, which exists in fetal liver but is lacking in vitro, turns iPSC-HEs into HSPCs. To test this, we transplanted CD45-negative iPSC-HEs into fetal sheep liver, in which HSPCs first grow. Within 2 months, the transplanted cells became CD45 positive and differentiated into multilineage blood cells in the fetal liver. Then, CD45-positive cells translocated to the bone marrow and were maintained there for 3 years with the capability of multilineage differentiation, indicating that hematopoietic cells with long-term engraftment potential were generated. Moreover, human hematopoietic cells were temporally enriched by xenogeneic donor-lymphocyte infusion into the sheep. This study could serve as a foundation to generate HSPCs from iPSCs.
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Experimental animals 69(2) 189-198 2019年12月 査読有り
MISC
10-
MOLECULAR THERAPY 25(5) 93-93 2017年5月
講演・口頭発表等
52-
Frontiers in Genome Engineering 2019 2019年11月26日
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Frontiers in Genome Engineering 2019 2019年11月26日
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第25回日本遺伝子細胞治療学会学術集会 2019年7月21日
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the 21st American Society of Gene and Cell therapy 2019年5月1日
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第41回日本分子生物学会 2018年11月30日 日本分子生物学会
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The 14th Nikko International Symposium 2017 2017年10月27日
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International Society for Stem Cell Research 2017 Annual Meeting 2017年6月16日
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the 20th American Society of Gene and Cell Therapy 2017年5月10日
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第15回 自治医科大学シンポジウム 2016年9月16日
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49th Annual Meeting of the Society for the Study of Reproduction 2016年7月
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49th Annual Meeting of the Society for the Study of Reproduction 2016年7月
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48th Annual Meeting of the Society for the Study of Reproduction 2015年6月
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40th Annucal Conference of the International Embryo Transfer Society 2014年1月
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The 6th International Conference on Advanced Fiber/Textile Materials 2011年12月
共同研究・競争的資金等の研究課題
4-
日本学術振興会 科学研究費補助金 若手研究 2019年4月 - 2021年3月
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リバネス リバネス研究費「メタジェン・腸内デザイン賞」 2018年4月 - 2019年3月
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日本学術振興会 科学研究費補助金 特別研究員奨励費 2013年4月 - 2015年3月
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日本学術振興会 科学研究費補助金 基盤研究(B) 2013年4月 - 2015年3月