Transgenesis in Hydra to characterize gene function and visualize cell behavior.


Journal

Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307

Informations de publication

Date de publication:
07 2019
Historique:
received: 02 10 2018
accepted: 04 04 2019
pubmed: 5 6 2019
medline: 26 11 2019
entrez: 5 6 2019
Statut: ppublish

Résumé

The freshwater polyp Hydra is a cnidarian used as a model organism in a number of fields, including the study of the origin and evolution of developmental mechanisms, aging, symbiosis and host-microbe interactions. Here, we describe a procedure for the establishment of stable transgenic Hydra lines by embryo microinjection. The three-stage protocol comprises (i) the design and preparation of a transgenic construct, (ii) the microinjection of the vector into early embryos of Hydra vulgaris, and (iii) the selection and enrichment of mosaic animals in order to develop uniformly transgenic clonal lines. The preparation of a transgenic construct requires ~2 weeks, and transgenic lines can be obtained within 3 months. The method allows constitutive or inducible gain- and loss-of-function approaches, as well as in vivo tracing of individual cells. Hydra polyps carrying transgenic cells reveal functional properties of the ancestral circuitry controlling animal development.

Identifiants

pubmed: 31160787
doi: 10.1038/s41596-019-0173-3
pii: 10.1038/s41596-019-0173-3
doi:

Substances chimiques

RNA, Small Interfering 0
Green Fluorescent Proteins 147336-22-9

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

2069-2090

Auteurs

Alexander Klimovich (A)

Zoological Institute, University of Kiel, Kiel, Germany. aklimovich@zoologie.uni-kiel.de.

Jörg Wittlieb (J)

Zoological Institute, University of Kiel, Kiel, Germany.

Thomas C G Bosch (TCG)

Zoological Institute, University of Kiel, Kiel, Germany. tbosch@zoologie.uni-kiel.de.

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Classifications MeSH