Hijacking oocyte DNA repair machinery in transgenesis?
Transgenesis refers to the modification of a genome to carry specified exogenous DNA sequences (transgenes, tgs) in a plant or animal; tgs are ideally transmissible through the germline. It has recently been shown that a membrane-disrupted spermatozoon mixed with exogenous DNA can be microinjected into an unfertilized, metaphase II (mII) oocyte to generate transgenic mouse embryos and offspring. This is here referred to as metaphase II (mII) transgenesis. Exogenous DNA clearly becomes genomically integrated in this process, but how? Presumably, the integration process utilizes DNA repair and recombinational machinery resident within mII oocytes. With recent advances in the description of DNA recombination and repair per se and in relation to meiotic and mitotic cell cycles, we are now poised to explain features of mIl transgenesis. Conversely, the method may of itself provide a new tool to probe these aspects of DNA metabolism. This article describes mil transgenesis in the context of DNA recombination and homeostasis in mII oocytes. A fuller understanding of the underlying recombinational mechanisms may enable improved methods of manipulating mammalian genomes and lead to gene targeting and genetic surgery in mII oocytes.
Perry
A C
AC
Laboratory of Developmental Biology and Neurogenetics, The Rockefeller University, New York, New York 10021, USA. perrya@rockvax.rockefeller.edu
D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000166:cytology; D004260:DNA Repair / Q000235:genetics*; D018816:Genes, cdc; D008677:Metaphase; D008845:Microinjections; D009865:Oocytes / Q000166:cytology*; D011995:Recombination, Genetic; D019076:Transgenes