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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
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Lysozyme Amyloid for Synthetic RNA Delivery.

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Lysozyme amyloid aggregates effectively deliver RNA, including immunostimulatory polyinosinic-polycytidylic acid (Poly(I:C)) and functional small interfering RNA (siRNA). This demonstrates their potential as safe and efficient non-viral RNA delivery vectors for immunotherapy applications.

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Area of Science:

  • Biomaterials Science
  • Nanotechnology
  • Immunology

Background:

  • Lysozyme-based amyloid aggregates possess favorable characteristics for RNA delivery, including stability, cationic charge, and biocompatibility.
  • These aggregates can form well-defined structures, making them suitable for drug carrier applications.
  • Their potential for delivering immunostimulatory synthetic RNA, such as polyinosinic-polycytidylic acid (Poly(I:C)), was investigated.

Purpose of the Study:

  • To evaluate lysozyme amyloid aggregates as non-viral drug carriers for RNA delivery.
  • To assess the efficacy of these aggregates in delivering Poly(I:C) and small interfering RNA (siRNA).
  • To confirm that observed effects were due to successful RNA delivery and not the complex itself.

Main Methods:

  • Characterization of lysozyme amyloid aggregates using particle size, zeta potential, and morphology analysis.
  • Assessment of RNA encapsulation efficiency.
  • In vitro studies using RAW 264.7 macrophage-like cells and A549-GFP cells to evaluate RNA uptake, immune activation, and gene silencing.

Main Results:

  • Poly(I:C)-loaded aggregates enhanced RNA uptake and induced significant immune activation in macrophage-like cells without toxicity.
  • GFP siRNA-loaded aggregates successfully delivered functional siRNA into A549-GFP cells, leading to a notable decrease in GFP expression.
  • Confocal microscopy and flow cytometry confirmed intracellular delivery and the functional impact of the delivered siRNA.

Conclusions:

  • Lysozyme amyloid aggregates show significant potential as effective non-viral vectors for RNA delivery.
  • These findings suggest promising applications in immunotherapy through targeted RNA delivery.
  • The study validates the use of lysozyme amyloids for delivering functional RNA payloads, including immunostimulatory and gene-silencing molecules.