Abstract
CRISPRs (clustered regularly interspaced short palindromic repeats), together with the nearby CRISPR-associated (cas) operon, constitute a prokaryotic RNA-based adaptive immune system against exogenous genetic elements. Here, we describe nuclease assays that are useful for characterizing the substrate-specific function of CRISPR-associated protein Cas2. We also provide methods for characterizing the stoichiometry and affinity between Cas2 and divalent metal ions using isothermal titration calorimetry (ITC).
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Acknowledgements
This work was supported by the National Institutes of Health Grant GM-102543 (to A.K.).
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Nam, K.H., DeLisa, M.P., Ke, A. (2015). Characterizing Metal-Dependent Nucleases of CRISPR-Cas Prokaryotic Adaptive Immunity Systems. In: Lundgren, M., Charpentier, E., Fineran, P. (eds) CRISPR. Methods in Molecular Biology, vol 1311. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2687-9_17
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DOI: https://doi.org/10.1007/978-1-4939-2687-9_17
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2686-2
Online ISBN: 978-1-4939-2687-9
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