Polyhistidine-Tag-Enabled Conjugation of Quantum Dots and Enzymes to DNA Nanostructures

Methods Mol Biol. 2022:2525:61-91. doi: 10.1007/978-1-0716-2473-9_6.

Abstract

DNA nanostructures self-assemble into almost any arbitrary architecture, and when combined with their capability to precisely position and orient dyes, nanoparticles, and biological moieties, the technology reaches its potential. We present a simple yet multifaceted conjugation strategy based on metal coordination by a multi-histidine peptide tag (Histag). The versatility of the Histag as a means to conjugate to DNA nanostructures is shown by using Histags to capture semiconductor quantum dots (QDs) with numerical and positional precision onto a DNA origami breadboard. Additionally, Histag-expressing enzymes, such as the bioluminescent luciferase, can also be captured to the DNA origami breadboard with similar precision. DNA nanostructure conjugation of the QDs or luciferase is confirmed through imaging and/or energy transfer to organic dyes integrated into the DNA nanostructure.

Keywords: Bioluminescence resonance energy transfer (BRET); DNA nanostructures; Histidine peptide tag (Histag); Luciferase, peptide nucleic acids; Quantum dots.

MeSH terms

  • Coloring Agents
  • DNA / chemistry
  • Histidine / chemistry
  • Luciferases / chemistry
  • Nanostructures*
  • Quantum Dots* / chemistry

Substances

  • Coloring Agents
  • polyhistidine
  • Histidine
  • DNA
  • Luciferases