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A DNA-transposon-based approach to functional screening in neural stem cells

Item Type:Article
Title:A DNA-transposon-based approach to functional screening in neural stem cells
Creators Name:Albieri, I. and Onorati, M. and Calabrese, G. and Moiana, A. and Biasci, D. and Badaloni, A. and Camnasio, S. and Spiliotopoulos, D. and Ivics, Z. and Cattaneo, E. and Consalez, G.G.
Abstract: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.
Keywords:Neural Stem Cells, Transposons, Screenings, Sleeping Beauty, Animals, Mice
Source:Journal of Biotechnology
ISSN:0168-1656
Publisher:Elsevier (The Netherlands)
Volume:150
Number:1
Page Range:11-21
Date:1 October 2010
Official Publication:https://doi.org/10.1016/j.jbiotec.2010.07.027
PubMed:View item in PubMed

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