Piggybacking functionalized DNA nanostructures into live-cell nuclei.
Golbarg M RoozbahaniPatricia ColosiAttila OraveczElena M SorokinaWolfgang G PfeiferSiamak ShokriYin WeiPascal DidierMarcello DeLucaGaurav AryaLàszlò ToraMelike LakadamyaliMichael G PoirierCarlos E CastroPublished in: Science advances (2024)
DNA origami nanostructures (DOs) are promising tools for applications including drug delivery, biosensing, detecting biomolecules, and probing chromatin substructures. Targeting these nanodevices to mammalian cell nuclei could provide impactful approaches for probing, visualizing, and controlling biomolecular processes within live cells. We present an approach to deliver DOs into live-cell nuclei. We show that these DOs do not undergo detectable structural degradation in cell culture media or cell extracts for 24 hours. To deliver DOs into the nuclei of human U2OS cells, we conjugated 30-nanometer DO nanorods with an antibody raised against a nuclear factor, specifically the largest subunit of RNA polymerase II (Pol II). We find that DOs remain structurally intact in cells for 24 hours, including inside the nucleus. We demonstrate that electroporated anti-Pol II antibody-conjugated DOs are piggybacked into nuclei and exhibit subdiffusive motion inside the nucleus. Our results establish interfacing DOs with a nuclear factor as an effective method to deliver nanodevices into live-cell nuclei.
Keyphrases
- nuclear factor
- induced apoptosis
- toll like receptor
- cell cycle arrest
- drug delivery
- single molecule
- single cell
- endothelial cells
- endoplasmic reticulum stress
- transcription factor
- gene expression
- oxidative stress
- circulating tumor
- signaling pathway
- cancer therapy
- dna damage
- immune response
- quantum dots
- inflammatory response
- high resolution
- liquid chromatography
- tandem mass spectrometry