フランス視覚研究所・ソルボンヌ大学Livet研究室の隈元拓馬博士をお招きしてセミナーを開催致します。神経発生研究のための神経細胞多色標識法およびイメージング技術についてご講演いただきます。ふるってご参加下さい。学部学生・大学院生の参加も歓迎です。セミナーは英語ですが、質疑は日本語でも英語でも可能です。
演者： 隈元 拓馬 博士
所属： Jean LIVET Lab, Institut de la Vision-Sorbonne Université, France
タイトル： A novel genome integrating vector system for direct transgenesis
in neural development
日時： 2019年12月5日（木）17:00～18:00
場所： 医学部基礎研究A棟1階 第2講義室（生協向かい）
Abstract: In developmental neuroscience, direct transfection of neural stem cells in vivo with genome-integrating vectors is increasingly used as an alternative to the generation of transgenic animals. However, current genome-integrative vectors present important limitations: retroviral vectors traditionally used in cell lineage studies are cumbersome to produce and limited in cargo capacity. Naked DNA vectors are considerably simpler to use and can accept large transgenes, but they enable expression from non-integrated episomes in addition to that of integrated transgenes. This creates a transient burst of expression that prevents reliable identification of transgenic cells and can cause transgene leakiness and toxicity. We bypass this issue with a novel expression strategy termed “iOn” (integration-coupled On expression) switch, enabling to activate DNA transgenes introduced by standard transfection procedures as they integrate into the host genome, while episomal transgenes remain silent. Toxic and non-physiological effects due to high episomal expression are thus avoided. Marker expression solely reflects the activity of integrated transgenes, enabling straightforward identification and analysis of integration events directly after transfection.
We demonstrate the efficiency of the iOn strategy for stable additive transgenesis in neural progenitors in the embryonic cortex, retina and spinal cord of mice and chicken. In these systems, we show that iOn vectors coupled with fluorescent color reporters make an ideal tool for cell lineage tracing and open the way to functional mosaic analysis through simple and rapid somatic transfection procedures. iOn vectors also drive leak-proof genome integration-dependent Cre-lox recombination, which we use to reveal the clonal output of a genetically identified subtype of retinal progenitors in the developing chick retina. We also show that iOn vectors provide an efficient means to stably transfect and rapidly isolate clones of stem/pluripotent cells in vitro. These results establish iOn as a highly efficient and versatile strategy for “direct transgenesis” in any model accessible to transfection.
世話人： 医学研究院・疾患情報研究分野 今井 猛
#第13回疾患情報研究分野主催（第17回今井研主催）公開セミナー