Abstract
CRISPR/Cas9-based genome editing is an inexpensive and efficient tool for genetic modification. Here we present a methodological approach of establishing interleukin-17 receptor B (IL17RB) knockout cell lines using CRISPR/Cas9-mediated genomic deletion. IL17RB gene encodes for a cytokine receptor that specifically binds to IL17B and IL17E and overexpressed in various cancers. The method involves CRISPR design, CRISPR cloning, delivery of CRISPR clone into cells, and verification of IL17RB gene deletion by deletion screening primer design, genomic DNA extraction, and polymerase chain reaction (PCR). Similar approaches can be used for generating mammalian cell lines with gene knockout for other genes of interest.
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Acknowledgment
This work was supported by St. John’s University and NIH grant CA213426 to Yan Zhu.
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Hu, O., Provvido, A., Zhu, Y. (2020). Generation of IL17RB Knockout Cell Lines Using CRISPR/Cas9-Based Genome Editing. In: Vancurova, I., Zhu, Y. (eds) Immune Mediators in Cancer. Methods in Molecular Biology, vol 2108. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0247-8_28
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DOI: https://doi.org/10.1007/978-1-0716-0247-8_28
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