Vitro and Ex Vivo Methods for Inducing Modifications of Target Endogenous Nucleic Acid Sequences in Nucleuses of Eukaryotic Cells

Abstract

The present disclosure relates to in vitro and ex vivo methods for inducing targeted mutagenesis of an endogenous DNA in a eukaryotic cell. Methods disclosed herein may comprise introducing a composition comprising a Cas9 protein and a guide RNA into a eukaryotic cell. In some embodiments, the Cas9 protein is complexed with the guide RNA to form an active endonuclease before being introduced into the eukaryotic cell. The guide RNA may be a dual guide RNA or a single guide RNA comprising a CRISPR RNA (crRNA) and a transactivating crRNA (tracrRNA). In some embodiments, the composition may be introduced into the cell by microinjection, electroporation, DEAE-dextran treatment, lipofection, nanoparticle-mediated transfection, or protein transduction domain mediated transduction.

Dependencies are inferred from the claim text.