A novel strategy combining array-CGH, whole-exome sequencing and in utero electroporation in rodents to identify causative genes for brain malformations

Valerio Conti, Aurelie Carabalona, Emilie Pallesi-Pocachard, Richard J. Leventer, Fabienne Schaller, Elena Parrini, Agathe A. Deparis, Françoise Watrin, Emmanuelle Buhler, Francesca Novara, Stefano Lise, Alistair T. Pagnamenta, Usha Kini, Jenny C. Taylor, Orsetta Zuffardi, Alfonso Represa, David Antony Keays, Renzo Guerrini, Antonio Falace, Carlos Cardoso

Research output: Contribution to journalArticle

Abstract

Birth defects that involve the cerebral cortex-also known as malformations of cortical development (MCD)-are important causes of intellectual disability and account for 20-40% of drug-resistant epilepsy in childhood. High-resolution brain imaging has facilitated in vivo identification of a large group of MCD phenotypes. Despite the advances in brain imaging, genomic analysis and generation of animal models, a straightforward workflow to systematically prioritize candidate genes and to test functional effects of putative mutations is missing. To overcome this problem, an experimental strategy enabling the identification of novel causative genes for MCD was developed and validated. This strategy is based on identifying candidate genomic regions or genes via array-CGH or whole-exome sequencing and characterizing the effects of their inactivation or of overexpression of specific mutations in developing rodent brains via in utero electroporation. This approach led to the identification of the C6orf70 gene, encoding for a putative vesicular protein, to the pathogenesis of periventricular nodular heterotopia, a MCD caused by defective neuronal migration.

Original languageEnglish
Article numbere53570
JournalJournal of Visualized Experiments
Volume2017
Issue number130
DOIs
Publication statusPublished - Dec 1 2017

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Keywords

  • Animal model
  • Array-CGH
  • In utero electroporation
  • Issue 130
  • Malformations of cortical development
  • Neuroscience
  • RNA interference
  • Whole-exome sequencing

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Medicine(all)

Cite this

Conti, V., Carabalona, A., Pallesi-Pocachard, E., Leventer, R. J., Schaller, F., Parrini, E., Deparis, A. A., Watrin, F., Buhler, E., Novara, F., Lise, S., Pagnamenta, A. T., Kini, U., Taylor, J. C., Zuffardi, O., Represa, A., Keays, D. A., Guerrini, R., Falace, A., & Cardoso, C. (2017). A novel strategy combining array-CGH, whole-exome sequencing and in utero electroporation in rodents to identify causative genes for brain malformations. Journal of Visualized Experiments, 2017(130), [e53570]. https://doi.org/10.3791/53570