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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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A splice-switch oligonucleotide loaded self-cleavable DNA nanogel.

Yuwei Hu1,2, Feng Chen1,2,3, Hongfang Lu1,2,4

  • 1Institute of Materials Research and Engineering (IMRE), Agency for Science Technology and Research (A*STAR), 2 Fusionopolis Way, Innovis #08-03, Singapore 138634, Republic of Singapore. ywhu@imre.a-star.edu.sg.

Chemical Communications (Cambridge, England)
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A novel DNA nanogel releases splice-switch oligonucleotides (SSOs) when exposed to acidic conditions. This self-cleavable system ensures SSO delivery in an unaltered state, paving the way for advanced therapeutic applications.

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Area of Science:

  • Biotechnology
  • Materials Science
  • Molecular Biology

Background:

  • DNA nanogels offer potential for controlled drug delivery.
  • Efficient and non-damaging release of therapeutic oligonucleotides is crucial.
  • Acid-labile linkers and i-motif structures can be utilized for triggered release.

Purpose of the Study:

  • To develop a self-cleavable DNA nanogel system for splice-switch oligonucleotide (SSO) delivery.
  • To investigate the release mechanism of SSO from the nanogel under acidic conditions.
  • To confirm the integrity of the released SSO.

Main Methods:

  • Fabrication of a DNA nanogel loaded with SSO.
  • Treatment of the nanogel with acidic buffer (pH 5.0).
  • Analysis of nanogel disintegration and SSO release using appropriate techniques.

Main Results:

  • The DNA nanogel successfully disintegrated under acidic conditions (pH 5.0).
  • Cleavage of the acid-labile linker initiated the i-motif structure formation.
  • Efficient release of intact, native splice-switch oligonucleotides was achieved.

Conclusions:

  • The developed self-cleavable DNA nanogel provides a robust platform for triggered SSO release.
  • Acidic environments can effectively trigger nanogel disassembly and SSO liberation.
  • This system preserves the structural integrity of the released SSO, suitable for therapeutic use.