Abstract
Base editors, such as cytosine and adenine base editors, are composed of nickase Cas9 (nCas9) and deaminase and serve as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based enzymatic tools for specific nucleotide substitutions. They are mainly the most effective genome editing tools for introducing point mutations, such as C-to-T and A-to-G conversions. The enhanced base editor, a C-to-G base editor (CGBE), can perform other nucleotide substitutions, such as C-to-G conversions. Here, we introduce a method for generating mouse models with point mutations using a base editing system.
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References
Chapman JR, Taylor MR, Boulton SJ (2012) Playing the end game: DNA double-strand break repair pathway choice. Mol Cell 47(4):497–510. https://doi.org/10.1016/j.molcel.2012.07.029
Anzalone AV, Koblan LW, Liu DR (2020) Genome editing with CRISPR-Cas nucleases, base editors, transposases and prime editors. Nat Biotechnol 38(7):824–844. https://doi.org/10.1038/s41587-020-0561-9
Komor AC, Kim YB, Packer MS, Zuris JA, Liu DR (2016) Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 533(7603):420–424. https://doi.org/10.1038/nature17946
Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable base editing of A*T to G*C in genomic DNA without DNA cleavage. Nature 551(7681):464–471. https://doi.org/10.1038/nature24644
Kurt IC, Zhou R, Iyer S, Garcia SP, Miller BR, Langner LM, Grunewald J, Joung JK (2021) CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells. Nat Biotechnol 39(1):41–46. https://doi.org/10.1038/s41587-020-0609-x
<Vasectomizing a Mouse.pdf>
Takeo T, Nakagata N (2015) Superovulation using the combined administration of inhibin antiserum and equine chorionic gonadotropin increases the number of ovulated oocytes in C57BL/6 female mice. PLoS One 10(5):e0128330. https://doi.org/10.1371/journal.pone.0128330
<Pronuclear microinjection and oviduct transfer procedures for transgenic mouse production.pdf>
Kim K, Ryu SM, Kim ST, Baek G, Kim D, Lim K, Chung E, Kim S, Kim JS (2017) Highly efficient RNA-guided base editing in mouse embryos. Nat Biotechnol 35(5):435–437. https://doi.org/10.1038/nbt.3816
Ryu SM, Koo T, Kim K, Lim K, Baek G, Kim ST, Kim HS, Kim DE, Lee H, Chung E, Kim JS (2018) Adenine base editing in mouse embryos and an adult mouse model of Duchenne muscular dystrophy. Nat Biotechnol 36(6):536–539. https://doi.org/10.1038/nbt.4148
Abe T, Inoue KI, Furuta Y, Kiyonari H (2020) Pronuclear microinjection during S-phase increases the efficiency of CRISPR-Cas9-assisted knockin of large DNA donors in mouse zygotes. Cell Rep 31(7):107653. https://doi.org/10.1016/j.celrep.2020.107653
Demayo JL, Wang J, Liang D, Zhang R, Demayo FJ (2012) Genetically engineered mice by pronuclear DNA microinjection. Curr Protoc Mouse Biol 2:245–262. https://doi.org/10.1002/9780470942390.mo110168
Acknowledgments
This work was supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) of Korea (NRF-2020M3A9D5A01082439 and NRF-2019R1A2C2087198). The illustrations were generated using BioRender.com.
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Jeong, T.Y., Lim, SY., Seong, J.K., Kim, K. (2023). Targeted Mutagenesis in Mice Using a Base Editor. In: Bae, S., Song, B. (eds) Base Editors. Methods in Molecular Biology, vol 2606. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2879-9_9
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DOI: https://doi.org/10.1007/978-1-0716-2879-9_9
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