tagPAINT: covalent labelling of genetically encoded protein tags for DNA-PAINT imaging

Daniel J Nieves; Geva Hilzenrat; Jason Tran; Zhengmin Yang; Hugh H MacRae; Matthew A B Baker; J Justin Gooding; Katharina Gaus

doi:10.1098/rsos.191268

tagPAINT: covalent labelling of genetically encoded protein tags for DNA-PAINT imaging

R Soc Open Sci. 2019 Dec 11;6(12):191268. doi: 10.1098/rsos.191268. eCollection 2019 Dec.

Authors

Daniel J Nieves^{1

2

3}, Geva Hilzenrat^{2

3}, Jason Tran^{2

3}, Zhengmin Yang^{2

3}, Hugh H MacRae^{2

3}, Matthew A B Baker^{4

5}, J Justin Gooding^{6

7}, Katharina Gaus^{2

3}

Affiliations

¹ Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT, UK.
² EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, NSW 2052, Australia.
³ ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, NSW 2052, Australia.
⁴ School of Biotechnology and Biomolecular Science, University of New South Wales, Sydney, NSW 2052, Australia.
⁵ Commonwealth Scientific and Industrial Research Organisation (CSIRO), Synthetic Biology Future Science Platform, Brisbane, Australia.
⁶ School of Chemistry, University of New South Wales, Sydney, NSW 2052, Australia.
⁷ Australian Centre for NanoMedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW 2052, Australia.

Abstract

Recently, DNA-PAINT single-molecule localization microscopy (SMLM) has shown great promise for quantitative imaging; however, labelling strategies thus far have relied on multivalent and affinity-based approaches. Here, the covalent labelling of expressed protein tags (SNAP tag and Halo tag) with single DNA-docking strands and application of SMLM via DNA-PAINT is demonstrated. tagPAINT is then used for T-cell receptor signalling proteins at the immune synapse as a proof of principle.

Keywords: DNA-PAINT imaging; covalent; monovalent; protein tags; superresolution.

Associated data

figshare/10.6084/m9.figshare.c.4764581