title: Controlling gene loss of function in newts with emphasis on lens regeneration.
abstract: Here we describe a protocol for gene loss of function during regeneration in newts, specifically applied to lens regeneration. Knockdown with the use of morpholinos can be achieved both in vitro and in vivo, depending on the experimental design. These methods achieve desirable levels of gene knockdown, and thus can be compared with methods developed for use in other animals, such as zebrafish. The technology has been applied to study molecular mechanisms during the process of lens regeneration by knocking down genes at specific stages and examining their effects on other genes and lens differentiation. The protocol can take a few days or up to 20 d to complete, depending on the duration of the experiment.
authors.affiliation: Department of Biology and Center for Tissue Regeneration and Engineering, University of Dayton, Dayton, Ohio, USA. panagiotis.tsonis@notes.udayton.edu
authors.forename: Panagiotis A
authors.initials: PA
authors.lastname: Tsonis
authors.forename: Tracy
authors.initials: T
authors.lastname: Haynes
authors.forename: Nobuyasu
authors.initials: N
authors.lastname: Maki
authors.forename: Kenta
authors.initials: K
authors.lastname: Nakamura
authors.forename: Martin Miguel
authors.initials: MM
authors.lastname: Casco-Robles
authors.forename: Shouta
authors.initials: S
authors.lastname: Yamada
authors.forename: Tomoya
authors.initials: T
authors.lastname: Miura
authors.forename: Chikafumi
authors.initials: C
authors.lastname: Chiba
authors.forename: Katia
authors.initials: K
authors.lastname: Del Rio-Tsonis
mesh_terms: D000818:Animals; D055785:Gene Knockdown Techniques; D007908:Lens, Crystalline / Q000502:physiology*; D023421:Models, Animal; D012038:Regeneration; D012455:Salamandridae / Q000235:genetics*