title: Morpholinos: studying gene function in the chick.
abstract: The use of morpholinos for perturbing gene function in the chick, Gallus gallus, has led to many important discoveries in developmental biology. This technology makes use of in vivo electroporation, which allows gain and loss of function in a temporally, and spatially controlled manner. Using this method, morpholinos can be transfected into embryonic tissues from early to late developmental stages. In this article, we describe the methods currently used in our laboratory to knock down gene function using morpholinos in vivo. We also detail how morpholinos are used to provide consistency of the results, and describe two protocols to visualise the morpholino after electroporation. In addition, we provide guidance on avoiding potential pitfalls, and suggestions for troubleshooting solutions. These revised techniques provide a practical starting point for investigating gene function in the chick.
authors.affiliation: Department of Craniofacial Development & Stem Cell Biology, King's College London, Guy's Tower Wing, Floor 27, London SE1 9RT, UK.
authors.forename: Anneliese
authors.initials: A
authors.lastname: Norris
authors.affiliation: Department of Craniofacial Development & Stem Cell Biology, King's College London, Guy's Tower Wing, Floor 27, London SE1 9RT, UK. Electronic address: andrea.streit@kcl.ac.uk.
authors.forename: Andrea
authors.initials: A
authors.lastname: Streit
mesh_terms: D000818:Animals; D002642:Chick Embryo; D015536:Down-Regulation; D018274:Electroporation / Q000295:instrumentation / Q000379:methods; D047108:Embryonic Development / Q000235:genetics; D055785:Gene Knockdown Techniques / Q000379:methods*; D060172:Morpholinos; D014162:Transfection / Q000379:methods
chemical_list: D060172:Morpholinos
keywords: Avian embryo; Gene knockdown; In vivo electroporation; Loss-of-function