Expanding the genetic toolkit in Xenopus: Approaches and opportunities for human disease modeling

• Published in: Developmental biology Vol. 426; no. 2; pp. 325 - 335
• Main Authors: Tandon, Panna, Conlon, Frank, Furlow, J. David, Horb, Marko E.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-06-2017
• Subjects: Animal Husbandry - organization & administration; Animals; Base Pairing; CRISPR-Cas; CRISPR-Cas Systems; Disease Models, Animal; Gene Editing - methods; Gene Knock-In Techniques; Gene Knockout Techniques; Genome; Human disease model; Humans; J strain; Knock-in; Laboratory Animal Science - organization & administration; Selective Breeding; TALENs; Tetraploidy; Transcription Activator-Like Effector Nucleases; Xenopus - genetics; Xenopus laevis; Xenopus laevis - genetics; Xenopus tropicalis; TALENs; Xenopus laevis; CRISPR-Cas; J strain; Xenopus tropicalis; Knock-in; Human disease model
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1016/j.ydbio.2016.04.009

The amphibian model Xenopus, has been used extensively over the past century to study multiple aspects of cell and developmental biology. Xenopus offers advantages of a non-mammalian system, including high fecundity, external development, and simple housing requirements, with additional advantages of large embryos, highly conserved developmental processes, and close evolutionary relationship to higher vertebrates. There are two main species of Xenopus used in biomedical research, Xenopus laevis and Xenopus tropicalis; the common perception is that both species are excellent models for embryological and cell biological studies, but only Xenopus tropicalis is useful as a genetic model. The recent completion of the Xenopus laevis genome sequence combined with implementation of genome editing tools, such as TALENs (transcription activator-like effector nucleases) and CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR associated nucleases), greatly facilitates the use of both Xenopus laevis and Xenopus tropicalis for understanding gene function in development and disease. In this paper, we review recent advances made in Xenopus laevis and Xenopus tropicalis with TALENs and CRISPR-Cas and discuss the various approaches that have been used to generate knockout and knock-in animals in both species. These advances show that both Xenopus species are useful for genetic approaches and in particular counters the notion that Xenopus laevis is not amenable to genetic manipulations.
•An overview of CRISPR-Cas and TALEN genome editing advances in Xenopus laevis and Xenopus tropicalis that have the potential to revolutionize this classical model system.•Availability of X. laevis genome sequence makes this allotetraploid species amenable to genome editing.•Genome editing in this system offers unique opportunities for modeling human disease.•Unique biological advantages of the organism provides clever strategies to avoid FO lethality.•Technological advancements allow knock-in approaches in Xenopus allow for precision base pair changes and in–frame tagging of proteins expressed from their endogenous loci.