2. Academic Validation
  3. Unconventional Lysine-Type Lipid Assemblies Enable Efficient Antisense Oligonucleotide Delivery with Distinct Structural Features

Unconventional Lysine-Type Lipid Assemblies Enable Efficient Antisense Oligonucleotide Delivery with Distinct Structural Features

  • Pharmaceutics. 2026 Feb 11;18(2):228. doi: 10.3390/pharmaceutics18020228.
Jieyan He 1 2,  Whitney Shatz-Binder 3,  Alexandra Robles 3,  Nanzhi Zang 4,  Wei Jia 4,  Sakura Sahai 5,  Matthew C Johnson 6,  Jing Li 1,  Chun-Wan Yen 4,  Shinji Takeoka 7
Affiliations

Affiliations

  • 1 Biochemical and Cellular Pharmacology, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
  • 2 Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo 162-8480, Japan.
  • 3 Pharmaceutical Development, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
  • 4 Synthetic Molecule Pharmaceutical Sciences, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
  • 5 Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, Tokyo 162-8480, Japan.
  • 6 Department of Structural Biology, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
  • 7 Advanced Research Institute of Science and Engineering, Waseda University, Tokyo 169-8555, Japan.
PMID: 41754970 DOI: 10.3390/pharmaceutics18020228
Abstract

Background/Objectives: Antisense Oligonucleotides (ASOs) hold great therapeutic potential due to their precise ability to modulate gene expression, particularly for treating genetic and neurological disorders. However, effective delivery of ASOs remains a major challenge. While most recent research focused on lipid nanoparticles (LNPs) as ASO carriers, alternative formulations, preparation methods and lipid compositions on delivery optimization are not fully explored. In this study, we investigated two types of formulations, lipoplexes (LPXes) and LNPs, prepared using lysine-type Cationic Lipids, K3C14 or K3C16, selected from a lysine-type lipid mini-library for their superior formulation stability and distinct cellular entry mechanisms. Methods: The physicochemical properties of the formulations were characterized using dynamic light scattering. Cytotoxicity was evaluated in spleen and liver cell lines. LPXes and LNPs were assessed for ASO delivery efficiency using an engineered HEK293 split-luciferase cell line, while immune response was evaluated in human peripheral blood mononuclear cells. Cryogenic electron microscopy (Cryo-EM) images were captured for structural analysis. Results: Lysine-type lipid mini-library screening identified lipids with either a hydrocarbon spacer K3 or C14 fatty acid tail exhibiting great stability and safety. Among the tested LPX and LNP formulations, the K3C16 lipoplex demonstrated ASO delivery efficiency and immune responses comparable to the benchmark SpikeVax LNP formulation. Notably, Cryo-EM imaging revealed novel structures that have not been reported previously; the K3C14 lipoplex formed a rouleaux-like structure, whereas the K3C16 lipoplex exhibited a lipid nanosheet-like structure, distinct from the conventional LNP structure. Conclusions: These results highlight the potential of an unconventional type of lipid assembly for efficient ASO delivery.

Keywords

ASO; LNP; cationic lipids; lipoplex; non-viral delivery system; structure.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-134541
    99.97%, Ionizable Amino Lipid