转座酶
生物
转座因子
神经干细胞
干细胞
克隆(Java方法)
胚胎干细胞
睡美人转座系统
细胞生物学
干细胞标记物
计算生物学
遗传学
分子生物学
作者
Ilaria Albieri,Marco Onorati,Giovanna Calabrese,Alessia Moiana,Daniele Biasci,Aurora Badaloni,Stefano Camnasio,Dimitrios Spiliotopoulos,Zoltán Ivics,Elena Cattaneo,G. Giacomo Consalez
标识
DOI:10.1016/j.jbiotec.2010.07.027
摘要
We describe the use of DNA transposons as tools for carrying out functional screenings in murine embryonic stem (ES) cell-derived neural stem (NS) cells. NS cells are a new type of stem cells featuring radial glial properties, that undergoes symmetric cell division for an indefinite number of passages, expanding as a monolayer. In this model, the previously unreported Sleeping Beauty transposase M3A achieves an optimal blend of clone generation efficiency and low redundancy of integrations per clone, compared to the SB100X Sleeping Beauty variant and to the piggyBac transposon. The technology described here makes it possible to randomly trap genes in the NS cell genome and modify their expression or tag them with fluorescent markers and selectable genes, allowing recombinant cells to be isolated and expanded clonally. This approach will facilitate the identification of novel determinants of stem cell biology and neural cell fate specification in NS cells.
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