Center for Bio/Molecular Science and Engineering, U.S. Naval Research Laboratory Code 6900, Washington, DC, USA.
National Research Council, Washington, DC, USA.
Methods Mol Biol. 2022;2525:61-91. doi: 10.1007/978-1-0716-2473-9_6.
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.
DNA 纳米结构可以自我组装成几乎任何任意的结构,并且当与它们精确定位和定向染料、纳米粒子和生物部分的能力结合时,该技术就会发挥出其潜力。我们提出了一种简单而多方面的基于金属配位的多组氨酸肽标签(Histag)的偶联策略。Histag 作为将 DNA 纳米结构偶联的一种手段具有多功能性,通过使用 Histag 将具有数字和位置精度的半导体量子点(QD)捕获到 DNA 折纸基板上。此外,表达酶的 Histag,如生物发光荧光素酶,也可以以类似的精度被捕获到 DNA 折纸基板上。通过将集成到 DNA 纳米结构中的有机染料的成像和/或能量转移来确认 QD 或荧光素酶与 DNA 纳米结构的偶联。