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