Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox
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Updated: Jul 23, 2026

Generation of Genetically Modified Mice through the Microinjection of Oocytes
Published on: June 15, 2017
Valerie Erhardt1, Chrystal M Snow1, Vishnu Hosur1
1The Jackson Laboratory for Mammalian Genetics, Bar Harbor, ME, USA.
Abstract:
Precise integration of DNA constructs greater than 3 kb into mouse zygotes is difficult. Here, we present a protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox. We describe steps for choosing mouse strains with preplaced attachment sites. We then detail procedures for microinjecting mouse zygotes with the plasmid donor DNA construct to generate transgenic mice by recombination-mediated cassette exchange. This protocol has the potential for application in exploring the functional implications of large structural variations in cancer. For complete details on the use and execution of this protocol, please refer to Low et al.1 and Hosur et al.2.
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