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

Klaus N. Lovendahl, Amanda N. Hayward, Wendy R Gordon

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)7030-7035
Number of pages6
JournalJournal of the American Chemical Society
Volume139
Issue number20
DOIs
StatePublished - May 24 2017

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DNA
Fusion reactions
Proteins
Endonucleases
Single-Stranded DNA
DNA sequences
Genes
Imaging techniques
Molecules
link protein
Gene Editing

Cite this

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

In: Journal of the American Chemical Society, Vol. 139, No. 20, 24.05.2017, p. 7030-7035.

Research output: Contribution to journalArticle

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