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Methods Mol Biol. 2015;1311:265-76. doi: 10.1007/978-1-4939-2687-9_17.

Characterizing Metal-Dependent Nucleases of CRISPR-Cas Prokaryotic Adaptive Immunity Systems.

Author information

1
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, 14853, USA.

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).

PMID:
25981479
DOI:
10.1007/978-1-4939-2687-9_17
[Indexed for MEDLINE]

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