A Multicassette Gateway Vector Set for High Throughput and Comparative Analyses in Ciona and Vertebrate Embryos

Roure A, Rothba ̈ cher U, Robin F, Kalmar E, Ferone G, et al (2007) A Multicassette Gateway Vector Set for High Throughput and Comparative Analyses in Ciona and Vertebrate Embryos. PLoS ONE 2(9): e916. doi:10.1371/journal.pone.0000916

A tort, je me plaignais hier soir auprès d’une collègue qui fêtait son départ à la retraite, qu’il n’y avait pas des beaux exemples de la participation française à la biologie des systèmes. Je retire mes plaintes en lisant le papier d’Agnès Roure et al. De la belle vectorologie de derrière les Eppendorf qui ajoute à la boîte à outils du biologiste moléculaire qui bosse en embryologie. La lecture sera un peu ardue, mais ça vaut la peine.

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Background. The past few years have seen a vast increase in the amount of genomic data available for a growing number of taxa, including sets of full length cDNA clones and cis-regulatory sequences. Large scale cross-species comparisons of protein function and cis-regulatory sequences may help to understand the emergence of specific traits during evolution.
Principal Findings. To facilitate such comparisons, we developed a Gateway compatible vector set, which can be used to systematically dissect cis-regulatory sequences, and overexpress wild type or tagged proteins in a variety of chordate systems. It was developed and first characterised in the embryos of the ascidian Ciona intestinalis, in which large scale analyses are easier to perform than in vertebrates, owing to the very efficient embryo electroporation protocol available in this organism. Its use was then extended to fish embryos and cultured mammalian cells.
Conclusion. This versatile vector set opens the way to the mid-to large-scale comparative analyses of protein function and cis-regulatory sequences across chordate evolution. A complete user manual is provided as supplemental material.