A DNA origami-based device for investigating DNA bending proteins by transmission electron microscopy
| Autor: | Ashwin Karthick Natarajan, Anton Kuzyk, Joonas Ryssy |
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| Přispěvatelé: | Department of Neuroscience and Biomedical Engineering, Aalto-yliopisto, Aalto University |
| Rok vydání: | 2023 |
| Předmět: | |
| Zdroj: | Nanoscale. 15:3212-3218 |
| ISSN: | 2040-3372 2040-3364 |
| Popis: | The DNA origami technique offers precise positioning of nanoscale objects with high accuracy. This has facilitated the development of DNA origami-based functional nanomechanical devices that enable the investigation of DNA–protein interactions at the single particle level. Herein, we used the DNA origami technique to fabricate a nanoscale device for studying DNA bending proteins. For a proof of concept, we used TATA-box binding protein (TBP) to evaluate our approach. Upon binding to the TATA box, TBP causes a bend to DNA of ∼90°. Our device translates this bending into an angular change that is readily observable with a conventional transmission electron microscope (TEM). Furthermore, we investigated the roles of transcription factor II A (TF(II)A) and transcription factor II B (TF(II)B). Our results indicate that TF(II)A introduces additional bending, whereas TF(II)B does not significantly alter the TBP–DNA structure. Our approach can be readily adopted to a wide range of DNA-bending proteins and will aid the development of DNA-origami-based devices tailored for the investigation of DNA–protein interactions. |
| Databáze: | OpenAIRE |
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