A large size-selective DNA nanopore with sensing applications.
Rasmus P ThomsenMette Galsgaard MalleAnders Hauge OkholmSwati KrishnanSøren S-R BohrRasmus Schøler SørensenOliver RiesStefan VogelFriedrich C SimmelNikos S HatzakisJørgen KjemsPublished in: Nature communications (2019)
Transmembrane nanostructures like ion channels and transporters perform key biological functions by controlling flow of molecules across lipid bilayers. Much work has gone into engineering artificial nanopores and applications in selective gating of molecules, label-free detection/sensing of biomolecules and DNA sequencing have shown promise. Here, we use DNA origami to create a synthetic 9 nm wide DNA nanopore, controlled by programmable, lipidated flaps and equipped with a size-selective gating system for the translocation of macromolecules. Successful assembly and insertion of the nanopore into lipid bilayers are validated by transmission electron microscopy (TEM), while selective translocation of cargo and the pore mechanosensitivity are studied using optical methods, including single-molecule, total internal reflection fluorescence (TIRF) microscopy. Size-specific cargo translocation and oligonucleotide-triggered opening of the pore are demonstrated showing that the DNA nanopore can function as a real-time detection system for external signals, offering potential for a variety of highly parallelized sensing applications.
Keyphrases
- single molecule
- label free
- atomic force microscopy
- living cells
- electron microscopy
- circulating tumor
- molecular dynamics simulations
- deep learning
- machine learning
- loop mediated isothermal amplification
- risk assessment
- single cell
- real time pcr
- mass spectrometry
- cell free
- big data
- artificial intelligence
- high throughput
- high speed
- climate change
- sensitive detection
- optical coherence tomography