High-Speed Super-Resolution Imaging Using Protein-Assisted DNA-PAINT

被引：44
作者：
Filius, Mike [1 ]
Cui, Tao Ju [1 ]
Ananth, Adithya N. [1 ,2 ]
Docter, Margreet W. [1 ]
Hegge, Jorrit W. [3 ]
van der Oost, John [3 ]
Joo, Chirlmin [1 ]
机构：
[1] Delft Univ Technol, Kavli Inst Nanosci, Dept BioNanoSci, NL-2629 HZ Delft, Netherlands
[2] Netherlands Org Appl Sci Res TNO, NL-2600 AD Delft, Netherlands
[3] Wageningen Univ, Lab Microbiol, NL-6703 HB Wageningen, Netherlands
来源：
NANO LETTERS | 2020年 / 20卷 / 04期
基金：
欧洲研究理事会;
关键词：
DNA-PAINT; super-resolution microscopy; Ago-PAINT; single-molecule FRET; Argonaute; DNA origami; LOCALIZATION MICROSCOPY; ARGONAUTE; BINDING; MICRORNAS;
D O I：
10.1021/acs.nanolett.9b04277
中图分类号：
O6 [化学];
学科分类号：
0703 ;
摘要：
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.
引用
收藏
页码：2264 / 2270
页数：7
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