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

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.