Gene transfer by electroporation into hemogenic endothelium in the avian embryo

Authors

  • Catalina Ana Rosselló,

    1. Department of Cardiovascular Developmental Biology, Centro Nacional de Investigaciones Cardiovasculares (CNIC) Carlos III, Madrid, Spain
    Search for more papers by this author
  • Miguel Torres

    Corresponding author
    1. Department of Cardiovascular Developmental Biology, Centro Nacional de Investigaciones Cardiovasculares (CNIC) Carlos III, Madrid, Spain
    • Centro Nacional de Investigaciones Cardiovasculares (CNIC) Carlos III, Melchor Fernández Almagro, 3 28029 Madrid, Spain
    Search for more papers by this author

Abstract

Hematopoiesis is the dynamic process whereby blood cells are continuously produced in an organism. Blood cell production is sustained by a population of self-renewing multipotent hematopoietic stem cells (HSCs) throughout the life of an organism. Cells with definitive HSC properties appear in the mid-gestation embryo as dense clusters of cells budding from the floor of the aorta, and that of the vitelline and umbilical arteries in the aorta-gonads-mesonephros region. Attempts to genetically modify the aortic floor from which these HSCs arise have been unsuccessful in the mouse, since the regulation of gene expression in the hemogenic endothelium is largely unknown. Here we report the implementation of gene transfer by electroporation into dorsal aortic endothelial cells in the chick embryo. This approach provides a quick and reproducible method of generating gain/loss-of-function models to investigate the function of genes involved in HSC birth. Developmental Dynamics 239:1748–1754, 2010. © 2010 Wiley-Liss, Inc.

Ancillary