Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox.
Document Type
Article
Publication Date
4-14-2024
Original Citation
Erhardt V,
Snow C,
Hosur V.
Protocol for large DNA transgenesis in mice using the Cas9+Bxb1 toolbox. STAR Protoc. 2024;5(2):103022
Keywords
JMG, SS1
JAX Source
STAR Protoc. 2024;5(2):103022
ISSN
2666-1667
PMID
38625797
DOI
https://doi.org/10.1016/j.xpro.2024.103022
Grant
Research reported in this publication was partially supported by the National Cancer Institute under award numbers R01CA265978 and P30CA034196
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.