Expression and Purification of pAG-MNase for CUT&RUN.

Cleavage Under Targets and Release Using Nuclease (CUT&RUN) is a chromatin profiling strategy that releases specific DNA-protein complexes from in situ chromatin to the supernatant. CUT&RUN employs the hybrid protein pAG-MNase, composed of IgG binding proteins A and G, followed by micrococcal nuclease. Here, I describe the expression and purification of recombinant pAG-MNase as well as the assessment of its enzymatic activity. Purified pAG-MNase is hyperactive and enables large-scale analyses.
(© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)