Using inositol as a biocompatible ligand for efficient transgene expression

Abstract

Transgene transfection techniques using cationic polymers such as polyethylenimines (PEIs) and PEI derivatives as gene vectors have shown efficacy, although they also have shortcomings. PEIs have decent DNA-binding capability and good cell internalization performance, but they cannot deliver gene payloads very efficiently to cell nuclei. In this study, three hyperbranched polyglycerol-polyethylenimine (PG6-PEI) polymers conjugated with myo-inositol (INO) molecules were developed. The three resulting PG6-PEI-INO polymers have an increased number of INO ligands per molecule. PG6-PEI-INO 1 had only 14 carboxymethyl INO (CMINO) units per molecule. PG6-PEI-INO 2 had approximately 130 CMINO units per molecule. PG6-PEI-INO 3 had as high as 415 CMINO units approximately. Mixing PG6-PEI-INO polymers with DNA produced compact nanocomposites. We then performed localization studies using fluorescent microscopy. As the number of conjugated inositol ligands increased in PG6-PEI-INO polymers, there was a corresponding increase in accumulation of the polymers within 293T cell nuclei. Transfection performed with spherical 293T cells yielded 82% of EGFP-positive cells when using PG6-PEI-INO 3 as the vehicle. Studies further revealed that extracellular adenosine triphosphate (eATP) can inhibit the transgene efficiency of PG6-PEI-INO polymers, as compared with PEI and PG6-PEI that were not conjugated with inositol. Our work unveiled the possibility of using inositol as an effective ligand for transgene expression.

Chemical Synthesis, Versatile Structures and Functions of Tailorable Adjuvants for Optimizing Oral Vaccination

“…In contrast, although gene delivery strategies based on cationic biomaterials, such as PLA-modified or glycerol-conjugated PEI branched molecules, have shown potential for the mammalian cell expression of target antigens. Their security for oral administration must still be approved. , The current DNA oral vaccines include those developed on the basis of intestinal probiotics, such as Lactococcus lactis. This strategy is deemed relatively safe compared to those designed for expression within immune cells, which showed mutation risks .…”

“…Their security for oral administration must still be approved. 140,141 The current DNA oral vaccines include those developed on the basis of intestinal probiotics, such as Lactococcus lactis. 142 This strategy is deemed relatively safe compared to those designed for expression within immune cells, which showed mutation risks.…”