DNA-based Rolling Motors with Biotin & Streptavidin Brakes Público

Abstract

Inspired by biological motors, scientists have produced various molecular machines as model systems including a wide array of DNA walkers. However, another type of machine, DNA rolling motors, are deemed to be favorable in molecular level detection due to their high fidelity and speed. These motors, which roll atop a surface, promise to be the most sensitive and processive DNA machines, but they have yet to be used to detect different proteins. Therefore, in the following experiments, these motors were designed to detect concentrations of streptavidin using biotin as the sensor, and the velocities of DNA particles were measured at different streptavidin concentrations. Following extensive trials, it was found that velocity is, in fact, inversely related to streptavidin concentration lending. This highly sensitive system for detecting streptavidin can be extended to other molecules. Therefore, it offers great possibilities, such as, clinical applications in testing for various diseases. Accurate detection of the presence of such biomolecules has become increasingly important in early diagnosis and, therefore, in increasing survival rates for diseases.

Table of Contents

Table of Contents

1.   Introduction ………………………………………………………………………………….....1

2.   Experimental Methods ………………………………………………………………......….15

      2.1. Thermal evaporation of gold films

      2.2. Implementation of wells with IBIDI

      2.3.  Fabrication of DNA and biotinylated anchors of varying proportions

      2.4. Backfilling of the surface

      2.5. Hybridization of RNA to DNA anchoring strands

      2.6. Synthesis of azide functionalized particles

      2.7. Production of high density DNA silica particles

      2.8. Synthesis of biotinylated silica particles

      2.9. Addition of varying concentrations of streptavidin to biotinylated particles

      2.10. Preparation of rolling solution and imaging

      2.11. Particle Trajectory Analysis

      2.12. Streptavidin Fluorescence Calibration

      2.13. Testing for non-specific binding of streptavidin using flow cytometry

3.   Results and Discussion ………………………………………………………………............23

4.   Conclusion and Future Directions ………………………………………………………….42

5.   References ………………………………………………………………………………….........45

 

Figures

Figure 1 ……………………………………………………………….........................................1

Figure 2 …………………………………………………………………......................................2

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Figure 4 …………………………………………………………………......................................4

Figure 5 …………………………………………………………………......................................5

Figure 6 …………………………………………………………………......................................6

Figure 7 …………………………………………………………………......................................9

Figure 8 ……………………………………………………………….......................................12

Figure 9 ……………………………………………………………….......................................24

Figure 10 …………………………………………………………….........................................27

Figure 11 ……………………………………………………………….....................................29

Figure 12 …………………………………………………………….........................................32

Figure 13 …………………………………………………………….........................................36

Figure 14 …………………………………………………………….........................................38

Figure 15 …………………………………………………………….........................................39

Figure 16 …………………………………………………………….........................................40

Figure 17 …………………………………………………………….........................................41

Figure 18 …………………………………………………………….........................................44

  

Tables

Table 1 ……………………………………………………………….........................................13

Table 2 ……………………………………………………………….........................................25

Table 3 …………………………………………………………................................................43

About this Honors Thesis

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School	Emory College
Department	Physics
Degree	B.A.
Submission	Honors Thesis
Language	English
Research Field	Chemistry, Analytical
Palavra-chave	DNA monowheel streptavidin biotin DNA-based rolling motors sensing
Committee Chair / Thesis Advisor	Salaita, Khalid, Emory University
Committee Members	Luis Boigues-Lopez, José, Emory University Brody, Jed, Emory University

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