Methods for generating DNA free genome edited plants and the delivery of pre-assembled ribonucleoprotein complexes (RNP)—such as the Cas9/gRNA RNP complex—for highly efficient genome editing of cells, in particular of intact plant cells and tissues, selection of the transfected cells and regeneration of whole plants thereof.

A formulation is provided for application to a host plant to reduce, inhibit or impair one or more of growth and development of the host plant. A method of inhibiting growth plant growth and development is also provided as a means of controlling weedy species. The method comprises: selecting a suitable gene for growth suppression in a target plant; identifying an at least one target site accessible to base pairing in the suitable gene; identifying an at least one divergent site in the at least one target site; designing a construct complementary to the at least one divergent site; adding an at least one RNAi inducer to the construct; and delivering the construct to the target plant.

The present invention provides a poly(alkylene oxide) polymer based size selective method for enriching nucleic acid molecules having a length below a cut-off value from a nucleic acid containing sample, the method comprising: (a) preparing a binding mixture comprising —the nucleic acid containing sample, —a poly(alkylene oxide) polymer and —a salt and binding nucleic acid molecules of different sizes to a solid phase which comprises a functional group, preferably carboxylated magnetic particles; (b) separating the solid phase with the bound nucleic acid molecules from the remaining sample; and (c) contacting the solid phase with the bound nucleic acid molecules at least once with an elution composition comprising a poly(alkylene oxide) polymer and a salt to selectively elute nucleic acid molecules having a length below the cut-off value from the solid phase while larger nucleic acid molecules having a length above the cut-off value remain bound to the solid phase, wherein the concentration (w/v) of the poly(alkylene oxide) polymer in the elution composition is lower than the concentration (w/v) of the poly(alkylene oxide) polymer in the binding mixture of (a); (d) separating the solid phase with the bound larger nucleic acid molecules from the eluted nucleic acid molecules; and (e) optionally further purifying the eluted nucleic acid molecules. The method is particularly useful for separating extracellular target nucleic acids by size.

A method of transforming a mitochondrion of a plant cell to express a nitrogenase enzyme includes exposing the plant cell to a mitochondrial-targeting nanocarrier polypeptide and one or more nucleic acids encoding the nitrogenase enzyme. The one or more genes encoding the nitrogenase enzyme can be one or more Klebsiella nif genes. The method can be used to generate plants which have the capability of fixing atmospheric elemental nitrogen.

A method for transforming a plant includes exposing a shoot apex of an axillary bud of a plant body, and introducing, into the shoot apex, a microparticle coated with at least one kind of nucleic acid.

A formulation is provided for application to a host plant to reduce, inhibit or impair one or more of growth and development of the host plant. A method of inhibiting growth plant growth and development is also provided as a means of controlling weedy species. The method comprises: selecting a suitable gene for growth suppression in a target plant; identifying an at least one target site accessible to base pairing in the suitable gene; identifying an at least one divergent site in the at least one target site; designing a construct complementary to the at least one divergent site; adding an at least one RNAi inducer to the construct; and delivering the construct to the target plant.

Disclosed herein are compositions and methods for effecting alterations at a defined location in the genome of a non-epidermal plant cell. Further disclosed are methods for providing plants having a modified phenotype or a modified genome.

Methods and apparatus for the reproducible, consistent and efficacious delivery of an HBV vaccine to a subject. The disclosure comprises apparatus for the controlled administration of the HBV vaccine through an orifice to the subject, a plurality of penetrating electrodes arranged with a predetermined spatial relationship relative to the orifice, and an electrical signal generator operatively connected to the electrodes.

The present invention discloses A METHOD TO PRODUCE PROTEIN IN PENICILLIUM AMAGASAKIENSE'S SLEEPING SPORES BY TRANSFORMATION OF SSRNA. The method includes three steps of culture of Penicillium amagasakiense and collection of spores, pretreatment of Penicillium anagasakiense spores, and electroporation of Penicillium anagasakiense spores by using HDEN method. In the present invention, non-germinated spores are used as a starting material for introduction of exogenous molecules. The exogenous protein coding single stranded RNA is introduced into the resting spores of Penicillium amagasakiense by employing the HDEN electrotransformation technique to express protein. The method of this invention is simple and fast, the effect is excellent, and the transformation rate reaches more than 90%.

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.