A DNA origami-based device for investigating DNA bending proteins by transmission electron microscopy

Ashwin Karthick Natarajan*, Joonas Ryssy, Anton Kuzyk*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

1 Citation (Scopus)
44 Downloads (Pure)

Abstract

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.
Original languageEnglish
Pages (from-to)3212-3218
Number of pages7
JournalNanoscale
Volume15
Issue number7
Early online date17 Jan 2023
DOIs
Publication statusPublished - 17 Jan 2023
MoE publication typeA1 Journal article-refereed

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