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    Biomaterials

    (A-25) Long-Term and Room-Temperature Stability of DNA-Inspired Janus-Based Nanopieces for Large mRNA Delivery

    Friday, October 13, 2023
    9:30 AM – 10:30 AM PDT
    Location: Exhibit Hall - Row A - Poster # 25

      Presenting Author(s)

    • Jin Zhai (she/her/hers)

      Ph.D. student
      University of Connecticut, United States

      • Co-Author(s)

    • Jinhyung Lee

      Ph.D. Student
      University of Connecticut
      Windham, United States

    • WZ

      Wuxia Zhang

      Post-doctorate
      University of Connecticut, United States

      • Primary Investigator(s)

    • YC

      Yupeng Chen

      Professor
      University of Connecticut, United States

    Introduction:: Messenger RNA (mRNA) delivery is promising in various biomedical applications because it is particularly suitable for protein expression, such as genome editing, to reduce off-target effects. Lipid nanoparticles (LNPs) have been extensively utilized as the delivery platform for mRNA, like COVID-19 vaccines. However, the poor stability of LNPs and crucially required storage condition is still major drawback. In order to store LNP-mRNA longer, freezing and lyophilization have been widely used. Nevertheless, the pressure arising from crystallization and vacuum dehydration may result in damage to macromolecules resulting in a decrease in nanoparticle stability. In this study, we developed a new class of delivery vehicles called Janus-based nanopieces (JBNp) based on non-covalent DNA-inspired Janus-based nanotubes (JBNt), which maintain stability over 8 weeks of storage at room temperature.

    Materials and Methods:: The JBNp/mRNA was assembled by mixing JBNt and eGFP mRNA in nuclease-free water, followed by sonicated with a sonicator. For the stability study, JBNp with eGFP mRNA was stored at room temperature for over 8 weeks. Then, JBNp with eGFP mRNA was transfected to C28/I2 cells for 72h. Further, the delivery efficiency was determined by flow cytometry. The absorbance of JBNp with eGFP mRNA was evaluated with Ultraviolet-visible spectrophotometer. The size and surface charge of JBNp with eGFP mRNA was measured with Dynamic Light Scattering.

    Results, Conclusions, and Discussions:: (Figure 1A) The self-assembled JBNt was sonicated with mRNA to prepare JBNp-mRNA. (Figure 1B) TEM image of nano rod-shape vehicles with mRNA, JBNp-mRNA. (Figure 1C) The confocal laser scanning microscope image of cell delivery of JBNp shows the JBNp successfully delivered eGFP mRNA into cells. (Figure 1D) The delivery efficiency showed JBNp effectively delivered mRNA to cells over 8 weeks. (Figure 1E) The size and PDI suggested JBNp-mRNA maintained stable over 8 weeks.

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