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

Ilaria Albieri, Marco Onorati, Giovanna Calabrese, Alessia Moiana, Daniele Biasci, Aurora Badaloni, Stefano Camnasio, Dimitrios Spiliotopoulos, Zoltán Ivics, Elena Cattaneo, G. Giacomo Consalez

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)11-21
Number of pages11
JournalJournal of Biotechnology
Volume150
Issue number1
DOIs
Publication statusPublished - Oct 1 2010

    Fingerprint

Keywords

  • Neural stem cells
  • Screenings
  • Sleeping beauty
  • Transposons

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Albieri, I., Onorati, M., Calabrese, G., Moiana, A., Biasci, D., Badaloni, A., Camnasio, S., Spiliotopoulos, D., Ivics, Z., Cattaneo, E., & Consalez, G. G. (2010). A DNA transposon-based approach to functional screening in neural stem cells. Journal of Biotechnology, 150(1), 11-21. https://doi.org/10.1016/j.jbiotec.2010.07.027