The present invention provides nucleic acid-loaded virus-like particles (VLPs) that can be produced in plants and are designed to deliver a nucleic acid into mammalian cells. Also provided are pharmaceutical compositions comprising the nucleic acid-loaded VLPs, and methods of making and using the nucleic acid-loaded VLPs.

The present disclosure relates to plant-based recombinant protein production systems and their methods of production and use. The plant-based recombinant protein production system is a vector comprising a 5 UTR and a 3 UTR, wherein the 3 UTR comprises at least one terminator selected from the group consisting of: EU, IEU, NbACT3, NbACT617, NbACT567, Pin2, BDB501, BDB282, NbHSP, NbHSPb, Rep, RbcS, SIR, SIR 5/3, SIR 3, AtHSP, 35S, RepA, NOS, TMV, TNVD, PEMV, and BYDV. In certain implementations, the vector comprises two terminators in the 3 UTR, where the two terminators are fused to form a double terminator. For example, the double terminator comprises two members selected from the group consisting of: EU, IEU, NbACT3, NbACT617, NbACT567, Pin2, BDB501, BDB282, NbHSP, NbHSPb, Rep, RbcS, SIR, SIR 5/3, SIR 3, AtHSP, 35S, RepA, NOS, TMV, TNVD, PEMV, and BYDV. In some aspects, the vector further comprises a chromatin scaffold/matrix attachment region (MAR) that is downstream of the terminators.

Improved plant transient expression systems using optimized geminiviral vectors that efficiently produce heteromultimeric proteins are described herein. Examples of high yields are shown herein, including two, three, or four fluorescent proteins coexpressed simultaneously. Various antibodies were produced using the optimized vectors with special focus given to the creation and production of a chimeric broadly neutralizing anti-flavivirus antibody. The variable regions of this murine antibody, 2A10G6, were codon optimized and fused to a human IgG1. Analysis of the chimeric antibody showed that it was efficiently expressed in plants, can be purified to near homogeneity by a simple one-step purification process, retains its ability to recognize the Zika virus envelope protein, and induce an immune response against Zika virus. Two other monoclonal antibodies were produced at similar levels. This technology is versatile tool for the production of a wide spectrum of pharmaceutical multi-protein complexes in a fast, powerful, and cost-effective way.

A method for simply constructing a two-component virus vector and related applications. The two-component plant viral vector is optimized, so that a two-component virus genome is placed in a single plasmid. Due to the use of the single plasmid, the activation, resuspension and mixing processes of auxiliary bacteria are avoided, the workload of an Agrobacterium experimental stage is simplified by half, the potential cross-contamination risk in the bacteria mixing process is avoided, and the virus infectivity is improved, so that the use of the two-component viral vector is greatly simplified.

A method for simply constructing a two-component virus vector and related applications. The two-component plant viral vector is optimized, so that a two-component virus genome is placed in a single plasmid. Due to the use of the single plasmid, the activation, resuspension and mixing processes of auxiliary bacteria are avoided, the workload of an Agrobacterium experimental stage is simplified by half, the potential cross-contamination risk in the bacteria mixing process is avoided, and the virus infectivity is improved, so that the use of the two-component viral vector is greatly simplified.

The disclosure provides modified plant viruses designed for delivering a nucleotide of interest into mammalian cells. The modified plant viruses include a plant virus nucleotide sequence (e.g. fragment) that is capable of transfecting a mammalian cell when that mammalian cell expresses a receptor for the modified plant virus. Accordingly, the disclosure also provides receptors for the modified plant viruses as well as methods of using the receptor or the modified plant virus and receptor.

The present disclosure relates to plant-based recombinant protein production systems and their methods of production and use. The plant-based recombinant protein production system is a vector comprising a 5 UTR and a 3 UTR, wherein 3 UTR comprises at least one terminator selected from the group consisting of: EU, IEU, NbACT3, NbACT617, NbACT567, Pin2, BDB501, BDB282, NbHSP, NbHSPb, Rep, RbcS, SIR, SIR 5/3, SIR 3, AtHSP, 35S, RepA, NOS, TMV, TNVD, PEMV, and BYDV. In certain implementations, the vector comprises two terminators in 3 UTR, where the two terminators are fused to form a double terminator. For example, the double terminator comprises two members selected from the group consisting of: EU, IEU, NbACT3, NbACT617, NbACT567, Pin2, BDB501, BDB282, NbHSP, NbHSPb, Rep, RbcS, SIR, SIR 5/3, SIR 3, AtHSP, 35S, RepA, NOS, TMV, TNVD, PEMV, and BYDV. In some aspects, the vector further comprises a chromatin scaffold/matrix attachment region (MAR) that is downstream of the terminators.