Composition for cleavage of a target DNA comprising a leader RNA specific for the target DNA and CAS PROTEIN-encoding nucleic acid or CAS protein, and its use

摘要

The invention provides a CRISPR/Cas9 composition for cleaving a target DNA in a plant cell, wherein the target DNA is endogenous DNA, and wherein the composition comprises (a) a nucleic acid encoding a CRISPR-associated protein 9 (Cas9) polypeptide, or a Cas9 polypeptide; and (b) a guide RNA specific to the target DNA, wherein the guide RNA is (i) a dual guide RNA comprising a CRISPR RNA (crRNA) and a trans activating crRNA (tracrRNA); or (ii) a single-chain guide RNA comprising a CRISPR RNA (crRNA) fused to a trans activating crRNA (tracrRNA). The invention further provides a plant cell, in particular a protoplast, and a plant, comprising the composition of the invention as well as a method for cleaving a target DNA or introducing two nicks into a target DNA of a plant cell using the composition of the invention.