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Xenopus laevis transgenesis by sperm nuclear injection

Smith, Stuart J., Fairclough, Lynne, Latinkic, Branko, Sparrow, Duncan B. and Mohun, Timothy J. 2006. Xenopus laevis transgenesis by sperm nuclear injection. Nature Protocols 1 (5) , pp. 2195-2203. 10.1038/nprot.2006.325

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Abstract

The stable integration of transgenes into embryos of the frog Xenopus laevis is achieved using the procedure described here. Linear DNA containing the transgene is incorporated randomly into sperm nuclei that have had their membranes disrupted with detergent treatment. Microinjection of these nuclei into unfertilized eggs produces viable embryos that can be screened for activity of the transgene. The proportion of embryos that harbor the transgene varies from 10 to 40% of the total number of surviving embryos. Multiple copies of the transgene can integrate as a concatemer into the sperm genome, and more than one site of DNA integration might occur within resulting animals. Germ cell transmission of the transgene is routine and the procedure is well suited to the production of transgenic reporter frog lines. One day should be allocated for the preparation of the sperm nuclei, which are stored as aliquots for future use. The transgenesis reaction and egg injection take one morning.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: Nature Publishing Group
ISSN: 1754-2189
Last Modified: 04 Jun 2017 06:40
URI: http://orca.cf.ac.uk/id/eprint/63426

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