A combination of transposable elements and magnetic cell sorting provides a very efficient transgenesis system for chicken primary erythroid progenitors.
2009
Mejia-Pous, Camila | Viñuelas, José | Koszela, Joanna | Kawakami, Koichi | Takahashi, Yoshiko | Gandrillon, Olivier | Faure, Claudine | Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC) ; Université Claude Bernard Lyon 1 (UCBL) ; Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS) | Biologie Fonctionnelle, Insectes et Interactions (BF2I) ; Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées de Lyon (INSA Lyon) ; Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA) | Department of Geophysics [Sendai] ; Tohoku University [Sendai] | LNCC, the CNRS, the UCBL, the Region Rhône Alpes, the IXXI and the ARC
International audience
Afficher plus [+] Moins [-]anglais. BACKGROUND: Stable transgenesis is an undeniable key to understanding any genetic system. Retrovirus-based insertional strategies, which feature several technical challenges when they are used, are often limited to one particular species, and even sometimes to a particular cell type as the infection depends on certain cellular receptors. A universal-like system, which would allow both stable transgene expression independent of the cell type and an efficient sorting of transfected cells, is required when handling cellular models that are incompatible with retroviral strategies. RESULTS: We report here on the combination of a stable insertional transgenesis technique, based on the Tol2 transposon system together with the magnetic cell sorting (MACS) technique, which allows specific selection of cells carrying the transgene in an efficient, reliable and rapid way. CONCLUSION: This new Tol2/MACS system leads to stable expression in a culture of primary chicken erythroid cells highly enriched in cells expressing the transgene of interest. This system could be used in a wide variety of vertebrate species.
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