Transgenesis procedures in Xenopus

Abstract

Stable integration of foreign DNA into the frog genome has been the purpose of several studies aimed at generating transgenic animals or producing mutations of endogenous genes. Inserting DNA into a host genome can be achieved in a number of ways. In Xenopus, different strategies have been developed which exhibit specific molecular and technical features. Although several of these technologies were also applied in various model organizms, the attributes of each method have rarely been experimentally compared. Investigators are thus confronted with a difficult choice to discriminate which method would be best suited for their applications. To gain better understanding, a transgenesis workshop was organized by the X-omics consortium. Three procedures were assessed side-by-side, and the results obtained are used to illustrate this review. In addition, a number of reagents and tools have been set up for the purpose of gene expression and functional gene analyses. This not only improves the status of Xenopus as a powerful model for developmental studies, but also renders it suitable for sophisticated genetic approaches. Twenty years after the first reported transgenic Xenopus, we review the state of the art of transgenic research, focusing on the new perspectives in performing genetic studies in this species.

Growth, Development, and Intestinal Remodeling Occurs in the Absence of Thyroid Hormone Receptor α in Tadpoles of Xenopus tropicalis

“…Importantly, mechanisms of TH signaling in gene regulation and development are highly conserved between frogs and humans, and TH-dependent stages in tadpoles are comparable to perinatal stages in humans (13). Thus, because frogs are easy to breed and maintain in the laboratory, are the closest vertebrate relatives to humans that have free-living larvae with readily manipulable endocrine signaling, and have all the modern tools of a genetic model system, including a sequenced genome, an ORFeome, transgenesis, and gene knockout technology (18)(19)(20)(21), amphibian development is a particularly compelling model system for use in elucidating the actions and mechanisms of TH signaling during development.…”

Myogenic waves and myogenic programs during Xenopus embryonic myogenesis

“…The restriction enzyme-mediated integration transgenesis protocol was carried out as described previously (Kroll and Amaya, 1996;Chesneau et al, 2008). Transgenic XMyf5-gfp embryos were obtained by breeding a transgenic male carrying a transgene of Xenopus tropicalis Myf5 promoter sequence driving expression of gfp (Polli and Amaya, 2002) with a wildtype female.…”

Optimization of gene delivery methods in Xenopus laevis kidney (A6) and Chinese hamster ovary (CHO) cell lines for heterologous expression of Xenopus inner ear genes

“…Xenopus affords novel opportunities for genetic investigations of inner ear structure and function due to the availability of a sequenced genome and established methods for the facile production of transgenic animals (Chesneau et al 2008; Hellsten et al 2010). The ability to characterize prevalent Xenopus inner ear genes through heterologous expression in cell lines expands the repertoire of available methods for genetic analysis beyond the traditional and well-established Xenopus oöcyte expression system (Terhag et al 2010).…”