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


ABSTRACT: 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.

SUBMITTER: Lovendahl KN 

PROVIDER: S-EPMC5517037 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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

Lovendahl Klaus N KN   Hayward Amanda N AN   Gordon Wendy R WR  

Journal of the American Chemical Society 20170516 20


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 demons  ...[more]

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