Bacterial Artificial Chromosome Transgenesis Through Pronuclear Injection of Fertilized Mouse Oocytes
Kristina Vintersten, Giuseppe Testa, Ronald Naumann, Konstantinos Anastassiadis, A. Francis Stewart
Abstract
Kristina Vintersten, Giuseppe Testa, Ronald Naumann, Konstantinos Anastassiadis, A. Francis Stewart
Abstract
In the mouse, conventional transgenes often produced unpredictable results mainly because they were too small to recapitulate a natural gene context. Bacterial artificial chromosomes (BACs) are large enough to encompass the natural context of most mammalian genes and consequently deliver more reliable recapitulations of their endogenous counterparts. Furthermore, recombineering methods now make it easy to engineer precise changes in a BAC transgene. Consequently, BACs have become the preferred vehicle for mouse transgenesis. Here, we detail methods for BAC transgenesis through pronuclear injection of fertilized oocytes.
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In the mouse, conventional transgenes often produced unpredictable results mainly because they were too small to recapitulate a natural gene context. Bacterial artificial chromosomes (BACs) are large enough to encompass the natural context of most mammalian genes and consequently deliver more reliable recapitulations of their endogenous counterparts. Furthermore, recombineering methods now make it easy to engineer precise changes in a BAC transgene. Consequently, BACs have become the preferred vehicle for mouse transgenesis. Here, we detail methods for BAC transgenesis through pronuclear injection of fertilized oocytes.
Key concepts: Transgenesis, Recombineering, Bacterial artificial chromosome, Biology, Context (archaeology), Transgene, Genetics, Chromosome