Abstract
Conditional alleles in genetically modified mice allow for the deletion of a gene of interest in a target tissue when combined with a tissue-specific Cre recombinase. A conditional allele is achieved by introducing LoxP sites around a critical exon, a gene, or a cluster of genes. Previously, conditional alleles were introduced in the mouse germline by classic gene targeting in embryonic stem cells, a challenging and time-consuming procedure. Now, conditional alleles can be generated directly in fertilized mouse eggs (zygotes) using the CRISPR/Cas9 technology. This one-step generation of mice is easier in design and faster. Here, we describe our achieved success rate, the considerations in design of a conditional allele, a detailed protocol to prepare the zygote injection mix, and the screening procedure to identify the new conditional knockout mouse strain.
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Pritchard, C.E.J., Kroese, L.J., Huijbers, I.J. (2017). Direct Generation of Conditional Alleles Using CRISPR/Cas9 in Mouse Zygotes. In: Eroshenko, N. (eds) Site-Specific Recombinases. Methods in Molecular Biology, vol 1642. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7169-5_2
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DOI: https://doi.org/10.1007/978-1-4939-7169-5_2
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7167-1
Online ISBN: 978-1-4939-7169-5
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