title: Generation of transgenic Hydra by embryo microinjection.
abstract: As a member of the phylum Cnidaria, the sister group to all bilaterians, Hydra can shed light on fundamental biological processes shared among multicellular animals. Hydra is used as a model for the study of regeneration, pattern formation, and stem cells. However, research efforts have been hampered by lack of a reliable method for gene perturbations to study molecular function. The development of transgenic methods has revitalized the study of Hydra biology(1). Transgenic Hydra allow for the tracking of live cells, sorting to yield pure cell populations for biochemical analysis, manipulation of gene function by knockdown and over-expression, and analysis of promoter function. Plasmid DNA injected into early stage embryos randomly integrates into the genome early in development. This results in hatchlings that express transgenes in patches of tissue in one or more of the three lineages (ectodermal epithelial, endodermal epithelial, or interstitial). The success rate of obtaining a hatchling with transgenic tissue is between 10% and 20%. Asexual propagation of the transgenic hatchling is used to establish a uniformly transgenic line in a particular lineage. Generating transgenic Hydra is surprisingly simple and robust, and here we describe a protocol that can be easily implemented at low cost.
authors.affiliation: Yale Stem Cell Center and Department of Cell Biology, Yale University School of Medicine; celina.juliano@yale.edu.
authors.forename: Celina E
authors.initials: CE
authors.lastname: Juliano
authors.affiliation: Yale Stem Cell Center and Department of Cell Biology, Yale University School of Medicine.
authors.forename: Haifan
authors.initials: H
authors.lastname: Lin
authors.affiliation: Department of Biological Chemistry and the Developmental Biology Center, University of California, Irvine; resteele@uci.edu.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Steele
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004247:DNA / Q000008:administration & dosage*; D005260:Female; D055785:Gene Knockdown Techniques; D006829:Hydra / Q000196:embryology / Q000235:genetics*; D008297:Male; D008845:Microinjections / Q000379:methods*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics
chemical_list: D004247:DNA