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Related Experiment Video

Updated: Jun 1, 2025

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Combinatorial Nanoparticle-Bound ssDNA Oligonucleotide Library Synthesized by Split-and-Pool Synthesis.

John V L Nguyen1, Ahlem Meziadi1, Christina Nassif2

  • 1Department of Biomedical Engineering, McGill University, 3775 University Street, Montreal, Quebec H3A 2B4, Canada.

ACS Applied Bio Materials
|January 20, 2025
PubMed
Summary

We developed a split-and-pool method to create synthetic single-stranded DNA (ssDNA) libraries on nanoparticles. This technique allows for scalable, controlled synthesis of diverse DNA sequences for applications like data storage and synthetic genomes.

Keywords:
DNA libraryDNA-functionalized nanoparticlesaptamermolecular engineeringnucleic acid engineeringoligonucleotide synthesis

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Area of Science:

  • Synthetic Biology
  • Nanotechnology
  • Oligonucleotide Synthesis

Background:

  • Synthetic single-stranded DNA (ssDNA) oligonucleotides are crucial for applications such as DNA aptamers, digital data storage, DNA origami, and synthetic genomes.
  • Precise control over ssDNA synthesis is vital for generating combinatorial sequences with user-defined parameters, requiring easy manipulation, sequence detection, and controlled elongation/termination.

Purpose of the Study:

  • To present a novel split-and-pool method for generating synthetic ssDNA oligonucleotides on nanoparticles.
  • To enable the creation of scalable combinatorial libraries of ssDNA with user-defined sequence parameters.

Main Methods:

  • Developed a split-and-pool strategy for ssDNA synthesis utilizing nanoparticles as solid supports.
  • Implemented a process involving DNA coupling to nanoparticles, ligation of double-digested fragments for orientation-specific synthesis, and attachment of a single-digested fragment for strand termination.

Main Results:

  • Successfully generated scalable, combinatorial nanoparticle-bound ssDNA libraries.
  • Demonstrated controllable strand lengths and confirmed the quality of both the synthesized DNA and the nanoparticle supports through characterization.

Conclusions:

  • The presented split-and-pool method provides a robust platform for producing diverse ssDNA libraries on nanoparticles.
  • This approach facilitates advancements in various fields requiring precisely synthesized ssDNA, including synthetic biology and information storage.