Super-resolution imaging allows for the visualization of cellular structures on a nanoscale level. DNA-PAINT (DNA point accumulation in nanoscale topology) is a super-resolution method that depends on the binding and unbinding of DNA imager strands. The current DNA-PAINT technique suffers from slow acquisition due to the low binding rate of the imager strands. Here we report on a method where imager strands are loaded into a protein, Argonaute (Ago), which allows for faster binding. Ago preorders the DNA imager strand into a helical conformation, allowing for 10 times faster target binding. Using a 2D DNA origami structure, we demonstrate that Ago-assisted DNA-PAINT (Ago-PAINT) can speed up the current DNA-PAINT technique by an order of magnitude, while maintaining the high spatial resolution. We envision this tool to be useful for super-resolution imaging and other techniques that rely on nucleic acid interactions.
Bibliographical noteFunding Information:
We thank Carolien Bastiaanssen, Sung Hyun Kim, and Bernd Rieger for critical reading and feedback. C.J. was supported by Vidi (864.14.002) of The Netherlands Organization for Scientific Research; an ERC Consolidator Grant (819299) of the European Research Council; and Vrije Programma (SMPS) of the Foundation for Fundamental Research on Matter. A.N.A. acknowledges TNO for seed ERP-Bio Nano programme for funding.
Copyright © 2020 American Chemical Society.
- DNA origami
- single-molecule FRET
- super-resolution microscopy