Method for producing genome-modified plants from plant protoplasts at high efficiency
AssigneesInstitute for Basic Science, Seoul National University R&Db Foundation, Aict
InventorsJin Soo Kim, Jungeun Kim, Sunghwa Choe, Je Wook Woo, Soon Il Kwon, Hyeran Kim
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Abstract
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.
Claims15
A method for increasing the production efficiency of a genome-edited plant from a plant protoplast, comprising the steps of:
- (i) editing GSL-ALK (Glucosinolateoxoglutarate-dependent dioxygenase homolog) gene of the plant protoplast by direct introduction of a Cas protein-guide RNA ribonucleoprotein (RNP) in which a Cas protein and a guide RNA in the form of naked RNA are pre-assembled into an isolated plant protoplast without using a vector;
- and
- (ii) producing a GSL-ALK-edited plant by regenerating the plant protoplast,
- wherein the guide RNA is selected from the group consisting of SEQ ID Nos. 12 to 18.
A method for producing a genome-edited Brassica oleracea plant from a plant protoplast of Brassica oleracea, comprising the steps of:
- (i) editing GSL-ALK (Glucosinolateoxoglutarate-dependent dioxygenase homolog) gene a genome of a plant the protoplast of Brassica oleracea by introducing a Cas9 protein-guide RNA ribonucleoprotein (RNP) in which a Cas9 protein and a guide RNA in the form of naked RNA are pre-assembled into an isolated plant protoplast of Brassica oleracea;
- and
- (ii) producing a GSL-ALK gene genome-edited Brassica oleracea plant by regenerating the plant protoplast of Brassica oleracea into a whole plant by forming a callus from the plant protoplast of Brassica oleracea and further culturing the callus;
- wherein the plant protoplast is derived from Brassica oleracea;
- and wherein the guide RNA is selected from the group consisting of SEQ ID Nos. 12 to 18.
Dependencies are inferred from the claim text.