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Massively Parallel Selection of NanoCluster Beacons.

Yu-An Kuo1, Cheulhee Jung2, Yu-An Chen1

  • 1Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, 78712, USA.

Advanced Materials (Deerfield Beach, Fla.)
|August 9, 2022
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Summary
This summary is machine-generated.

This study introduces a high-throughput method to discover novel polar opposite twins (POTs) in NanoCluster Beacons (NCBs). The findings reveal key DNA activator sequences for generating bright, multicolor silver nanocluster probes.

Keywords:
NanoCluster Beaconsfluorescent nanomaterialshigh-throughput screeningnext-generation sequencingsilver nanoclusters

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Area of Science:

  • Nanotechnology
  • Biochemistry
  • Molecular Biology

Background:

  • NanoCluster Beacons (NCBs) are multicolor silver nanocluster probes activated by DNA strands.
  • Discovering activators for specific NCB fluorescence, especially polar opposite twins (POTs), is challenging with low-throughput methods.

Purpose of the Study:

  • To develop a high-throughput screening method for identifying novel POT-NCBs.
  • To elucidate the role of specific DNA activator sequences in NCB fluorescence.
  • To establish a predictive model for designing multicolor NCBs.

Main Methods:

  • Utilized repurposed next-generation sequencing chips for high-throughput screening of approximately 40,000 activator sequences.
  • Analyzed fluorescence activation outcomes based on single-nucleotide variations in DNA activators.
  • Developed a
  • zipper-bag
  • model to explain observed phenomena.

Main Results:

  • Identified nucleobases at positions 7-12 of the 18-nucleotide activator as critical for bright NCB fluorescence.
  • Determined positions 4-6 and 2-4 as hotspots for generating yellow-orange and red POTs, respectively.
  • Established a correlation between activator sequence, silver cluster chromophore formation, and chemical yield.

Conclusions:

  • A high-throughput selection method significantly accelerates the discovery of POT-NCBs.
  • Specific DNA sequence motifs dictate NCB fluorescence properties and POT generation.
  • Integrated high-throughput screening with machine learning enables in silico design of bright, multicolor NCBs.