3 years ago

Sequence-Directed Covalent Protein–DNA Linkages in a Single Step Using HUH-Tags

Sequence-Directed Covalent Protein–DNA Linkages in a Single Step Using HUH-Tags
Klaus N. Lovendahl, Wendy R. Gordon, Amanda N. Hayward
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.

Publisher URL: http://dx.doi.org/10.1021/jacs.7b02572

DOI: 10.1021/jacs.7b02572

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