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Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Sequence-Directed Covalent Protein-DNA Linkages in a Single Step Using HUH-Tags.
Journal of the American Chemical Society 2017 May 25
We present a robust strategy to covalently link proteins and DNA using HUH-endonuclease domains as fusion partners (HUH-tags). We show that HUH-tags react robustly with specific sequences of unmodified single-stranded DNA, and we have identified five tags that react orthogonally with distinct DNA sequences. We demonstrate the versatility of HUH-tags as fusion partners in Cas9-mediated gene editing and the construction of doubly DNA-tethered proteins for single-molecule studies. Finally we demonstrate application to cellular imaging in live and fixed cells.
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