Abstract
The CRISPR/Cas9 system, a defense mechanism naturally occurring in prokaryotes, has been recently repurposed as an RNA-guided DNA targeting platform and widely used as a powerful tool for genome editing. Here we describe how to modify the carboxy-terminal region, called Fragment crystallizable (Fc) region, of a murine monoclonal antibody by replacing the heavy chain constant exons with those from a teleost fish antibody by the CRISPR/Cas9 system. We outline optimal conditions for knockout and knockin mechanisms to edit the Immunoglobulin heavy chain (IgH) constant region gene locus in a murine hybridoma cell line. A chimeric mouse–fish monoclonal antibody can be successfully produced by hybridoma cell lines engineered according to this protocol.
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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Ametrano, A., Coscia, M.R. (2022). Production of a Chimeric Mouse–Fish Monoclonal Antibody by the CRISPR/Cas9 Technology. In: Verde, C., Giordano, D. (eds) Marine Genomics. Methods in Molecular Biology, vol 2498. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2313-8_19
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DOI: https://doi.org/10.1007/978-1-0716-2313-8_19
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2312-1
Online ISBN: 978-1-0716-2313-8
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