Xenopus laevis transgenesis by sperm nuclear injection.
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.
Smith
Stuart J
SJ
Division of Developmental Biology, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Fairclough
Lynne
L
Latinkic
Branko V
BV
Sparrow
Duncan B
DB
Mohun
Timothy J
TJ
D000818:Animals; D030801:Animals, Genetically Modified; D002467:Cell Nucleus; D004625:Embryo, Nonmammalian; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods*; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D014982:Xenopus laevis / Q000196:embryology / Q000235:genetics*