Bacterial artificial chromosome transgenesis through pronuclear injection of fertilized mouse oocytes.
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.
Vintersten
Kristina
K
Mount Sinai Hospital, Samuel Lunenfeld Research Institute, Toronto, Ontario, Canada.
Testa
Giuseppe
G
Naumann
Ronald
R
Anastassiadis
Konstantinos
K
Stewart
A Francis
AF
D000777:Anesthetics; D000818:Animals; D002021:Buffers; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D004210:Dissection; D004624:Embryo Transfer; D005260:Female; D005306:Fertilization / Q000187:drug effects; D018014:Gene Transfer Techniques; D006728:Hormones / Q000008:administration & dosage / Q000494:pharmacology; D051379:Mice; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D010057:Oviducts / Q000187:drug effects; D010060:Ovulation / Q000187:drug effects; D019076:Transgenes / Q000235:genetics; D015053:Zygote / Q000378:metabolism*; D017388:Zygote Intrafallopian Transfer
D000777:Anesthetics; D002021:Buffers; D006728:Hormones; D004247:DNA