Preparation of siRNA encapsulated nanoliposomes suitable for siRNA delivery by simply discontinuous mixing

Abstract

Previously a scalable and extrusion-free method has been developed for efficient liposomal encapsulation of DNA by twice stepwise mixing of lipids in ethanol and DNA solution using T-shape mixing chamber. In this study, we prepared nanoliposomes encapsulating siRNA by simply discontinuous mixing of lipids in ethanol/ether/water mixture and acidic siRNA solution without use of special equipment. The simple mixing siRNA/liposomal particles (siRNA/SMLs) prepared using ethanol/ether/water (3:1:1) mixture showed 120.4 ± 20.2 nm particle size, 0.174 ± 0.033 polydispersity and 86.5 ± 2.76% siRNA encapsulation rate. In addition, the SMLs almost completely protected the encapsulated siRNA from RNase A digestion. Coupling of anti-human epidermal growth factor receptor (EGFR) Fab' to siRNA/SMLs enhanced EGFR-specific cell penetration of SMLs and induced siRNA dependent gene silencing. Unexpectedly, the Cy5.5-labeled Fab' showed almost no in vivo targeting to the xenografted A549 tumors in SCID-NOD mice. However, multiple injection of the unmodified siRNA/SMLs accumulated in the tumors and induced siRNA-dependent in vivo gene silencing. These results demonstrate that the siRNA/SMLs can be used as a siRNA delivery tool for gene therapy.

One-step microfluidics production of enzyme-loaded liposomes for the treatment of inflammatory diseases

“…For example, the encapsulation of biopharmaceuticals in liposomes leads to a modification in their pharmacokinetics and pharmacodynamics, improving their therapeutic activity [11]. Mokhtarieh et al have reported that the encapsulation of siRNA in liposomes prevented it from the RNase degradation and elimination [15]. Another example is related with the Cu,Zn-Superoxide dismutase (SOD).…”

The potential of siRNA based drug delivery in respiratory disorders: Recent advances and progress

“…The main difficulties of utilizing lipid‐based conveyance vectors in the clinical setup are their harmfulness and their nonspecific activation of inflammatory cytokines and interferon reactions. Lipid‐based delivery vectors can be classified into five types of molecules: Cationic lipoplexes and liposomes; PEGylated lipids; Neutral lipids; Lipids particles; and Lipid‐like molecules (Kaczmarek et al, ; Liu et al, ; Mokhtarieh, Lee, Kim, & Lee, ; Muddineti, Shah, Rompicharla, Ghosh, & Biswas, ).…”

“…The lipid-based delivery system is generally used to deliver siRNA in vitro and in vivo. Ordinarily, cationic lipids or liposomes utilizes negatively charged siRNA through electrostatic forces and are known as (Kaczmarek et al, 2018;Liu et al, 2019;Mokhtarieh, Lee, Kim, & Lee, 2018;Muddineti, Shah, Rompicharla, Ghosh, & Biswas, 2018).…”

Synchronous conjugation of i-motif DNA and therapeutic siRNA on the vertexes of tetrahedral DNA nanocages for efficient gene silence

“…There have been several studies reported that proteins 7 and nucleic acids 26 can be loaded in DNA nanocages by far. However, nucleic acids, like siRNA have to be delivered to the cytoplasm or nucleus to exert therapeutic effects 22 , 27 . In other words, lysosomal escape is one of the key processes for the intracellular disposal of nucleic acid drugs 28 , 29 .…”