Manipulation of Gene Function in Xenopus laevis
Xenopus laevis embryos are particularly well suited to address questions requiring either knockdown or overexpression of genes in a tissue-specific fashion during vertebrate embryonic development. These manipulations are achieved by targeted injection of either antisense morpholino oligonucleotides or synthetic mRNAs, respectively, into the early embryo. Herein we offer detailed protocols describing how to design and perform these experiments successfully, as well as a brief discussion of considerations for performing a microarray analysis in this organism.
Key wordsXenopus laevis embryogenesis microinjection morpholinos gene knockdown microarray
This work was supported by grants from the NIH (RO3HD058841 and RO1HD37976) and Shriners’ Hospital Research Foundation to JLC. MM was supported by NIH grant T32HD049309.
- 10.Maeno, M., Ong, R. C., Suzuki, A., Ueno, N., and Kung, H. F. (1994) A truncated bone morphogenetic protein 4 receptor alters the fate of ventral mesoderm to dorsal mesoderm: roles of animal pole tissue in the development of ventral mesoderm. Proc. Natl. Acad. Sci. USA 91, 10260–10264.PubMedCrossRefGoogle Scholar
- 19.Nieuwkoop, P. D. and Faber, J., and Hubrecht-Laboratory (1967) Normal table of Xenopus laevis (Daudin). A systematical and chronological survey of the development from the fertilized egg till the end of metamorphosis, 2 ed. North-Holland Pub. Co., Amsterdam.Google Scholar
- 20.Sive, H. L., Grainger, R. M., and Harland, R. M. (2000) Early Development of Xenopus laevis: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar