The present disclosure relates to complexes and compositions of cationic compounds combined with low mass percentages of PEGylated compounds, such as PEGylated lipids, for the delivery of nucleic acids and other polyanionic cargoes to cells, methods for preparing complexes and compositions comprising one or more cationic compounds and a low mass percentage of PEGylated compounds with polyanionic compounds, and methods for delivering the polyanionic compounds to cells.

The present invention is directed to improved methods for conjugating polyethylene glycol to polyethyleneimine, and to the use of such polyethylene glycol—polyethyleneimine conjugates to improve the efficiency with which viral and non-viral nucleic acid vectors transfect cells to provide gene therapy, and to improve the efficiency of producing viral particles that comprise therapeutic polynucleotides for use in gene therapy.

Disclosed herein are circular RNAs and transfer vehicles, along with related compositions and methods of treatment. The circular RNAs can comprise group I intron fragments, spacers, an IRES, duplex forming regions, and/or an expression sequence, thereby having the features of improved expression, functional stability, low immunogenicity, ease of manufacturing, and/or extended half-life compared to linear RNA. Pharmaceutical compositions comprising such circular RNAs and transfer vehicles are particularly suitable for efficient protein expression in immune cells in vivo. Also disclosed are precursor RNAs and materials useful in producing the precursor or circular RNAs, which have improved circularization efficiency and/or are compatible with effective circular RNA purification methods.

A Hepatitis E virus (HEV)-based virus like nanoparticle (HEVNP) made with a modified capsid protein containing at least a portion of open reading frame 2 (ORF2) protein conjugated with gold nanocluster is provided. Also provided are methods of targeted delivery of a nucleic acid using the HEVNP.

Disclosed herein are compositions and methods related to megakaryocyte-derived extracellular vesicles derived from human pluripotent stem cells, where the megakaryocyte-derived extracellular vesicles may be utilized for drug delivery and treating various diseases.

Described herein are muscle-specific targeting moieties and compositions including the muscle specific targeting motifs. Also described herein are uses of the muscle-specific targeting motifs and compositions including the muscle specific targeting moieties. In some embodiments, the muscle-specific targeting moieties and compositions including the muscle specific targeting moieties can be used to direct delivery of a cargo to a muscle cell.

RNA encoding an immunogen is delivered to a large mammal at a dose of between 2 μg and 100 μg. Thus the invention provides a method of raising an immune response in a large mammal, comprising administering to the mammal a dose of between 2 μg and 100 μg of immunogen-encoding RNA. Similarly, RNA encoding an immunogen can be delivered to a large mammal at a dose of 3 ng/kg to 150 ng/kg. The delivered RNA can elicit an immune response in the large mammal.

Provided are a method for preparing an mRNA-GalNAc targeting molecule, an in vivo delivery system therefor and use thereof. The mRNA-GalNAc targeting molecule comprises an mRNA molecule that is linked to PolyA modified with an N-acetylgalactosamine at 3′-end, wherein a sequence of the mRNA molecule comprises a 5′ cap and a target gene sequence. By directly linking the mRNA molecule expressing the target gene to the polyA sequence coupled with GalNAc, an mRNA molecule with GalNAc at 3′-end is synthesized to realize the aim of targeted drug delivery in liver. The method is simpler and more reliable, and solves existing problems in coupling between mRNA and N-acetylgalactosamine.

The disclosure provides methods and compositions for delivering RNA constructs to cells for functional expression and/or activity. In some aspects, the disclosure provides a composition comprising a multi-functionalized nanoparticle. The multi-functionalized nanoparticles comprise a core functionalized with at least one RNA molecule, at least one cell penetrating peptide (CPP), and at least one positively charged moiety, each of which is independently attached to the core, optionally with linker moieties. In some embodiments, the RNA molecule is an uncapped mRNA molecule with the 5′ end attached to a linker moiety that is attached to the core. The multi-functionalized nanoparticle is substantially neutral, negatively or positively charged. The multi-functionalized nanoparticle can be used in methods of delivering and causing the expression of polypeptides of interest in a cell for various purposes, including vaccination, cancer treatment, extension of telomeres, modification of cellular signaling pathways, and the like.

The present invention generally relates to composition matters and methods useful for gene delivery and an option for therapeutic treatment of various diseases. Particularly, this disclosure relates to a plasmid vector comprising a fusion of a plurality of genes comprising a gene of a chemokine or a cytokine, a gene for a targeting polypeptide and genes for one or more polypeptide linkers. Methods of use and composition matters are within the scope of this disclosure.