The present invention relates to a method of increasing the production efficiency of gene-edited plants, regenerated from plant protoplasts, by use of a Cas protein-guide RNA ribonucleoprotein (RNP). According to the present invention, the method of increasing the production efficiency of gene-edited plants makes it possible to efficiently produce target gene-mutated plants and to minimize the introduction of foreign DNA into plants. Thus, the present invention can be very advantageously used in a wide variety of fields, including agriculture, food and biotechnology.

The present invention relates to method for preparing a plant from a protoplast comprising knocking-out or knocking-in one or more the endogenous gene of the protoplast, and the plant regenerated from a genome-modified protoplast prepared by the above method.

A method includes introducing a protein having double-stranded DNA cleavage activity itself into the eukaryote or a part of the eukaryote, and rearranging DNA of the eukaryote by the protein in the eukaryote or a part of cells of the eukaryote.

Disclosed herein are plant-modifying compositions including a plurality of plant messenger packs (PMPs) (e.g., including a plant extracellular vesicle (EV), or segment, portion, or extract thereof), wherein the PMPs include a plant-modifying agent, and wherein the plant-modifying agent comprising the heterologous RNA. The compositions herein are useful in methods for modifying plants such as to increase plant fitness, wherein the increase in plant fitness comprising an increase in development, growth, yield, resistance to abiotic stressors, or resistance to biotic stressors.

Disclosed herein is a vaccine comprising an antigen and IL-33. Also disclosed herein is a method for increasing an immune response in a subject in need thereof. Further disclosed herein is a method for treating cancer in a subject in need thereof. The methods may comprise administering the vaccine to the subject.

Method for targeted alteration of a duplex acceptor DNA sequence in a plant cell protoplast, comprising combining the duplex acceptor DNA sequence with a donor mutagenic nucleobase, wherein the duplex acceptor DNA sequence contains a first DNA sequence and a second DNA sequence which is the complement of the first DNA sequence and wherein the donor mutagenic nucleobase comprises at least one mismatch with respect to the duplex acceptor DNA sequence to be altered, preferably with respect to the first DNA sequence, wherein the method further comprises a step of introducing the donor mutagenic nucleobase into the cell protoplasts using polyethylene glycol (PEG) mediated transformation and the use of PEG protoplast transformation for enhancing the rate of targeted mutagenesis.

The invention provides novel methods and compositions for transfer of nuclear and/or plastomic genomes, or portions thereof, or cytoplasmic component(s) and/or genetic material, between plant cells. Methods for production of a wounded mixed cell culture, or mixing two or more cell cultures after wounding, and transfer of genetic and/or cytoplasmic component(s), such as transfer of nuclear and/or plastid gene(s) or mutations, edits or alleles, between cells of the mixed culture, are also provided. Wounded mixed cell cultures produced by such methods, and resulting cells and regenerated plants, plant parts, and progeny plants are further provided. Molecular and genetic analyses, and screenable and selection markers, are also provided to confirm transfer and presence of cytoplasmic and/or nuclear component(s) and/or gene(s), mutation(s) or allele(s) in cells and plants produced by these methods.

The invention provides novel methods for liquid-mediated delivery of pollen grains to enclosed stigmas in recipient female flowers. For example, methods for liquid-mediated pollination are provided. The methods provided include collecting pollen from a donor plant, suspending the collected pollen in a liquid solution, and introducing said solution to an enclosed stigma of a recipient flower bud on a recipient plant, thereby pollinating the flower with the pollen from the donor plant.

A nanomaterial conjugate for delivering an active agent to a plant comprising a nanocarrier and an active agent linked to the nanocarrier. The nanocarrier is cellulose nanocrystal (CNC), and the nanocarrier delivers the active agent across cell walls of cells of the plant.

A method of introducing naked dsRNA into a seed is provided. The method comprising contacting the seed with the naked dsRNA under conditions which allow penetration of the dsRNA into the seed, thereby introducing the dsRNA into the seed.