Methods for Modifying Endogenous Nucleic Acid Sequences via Electroporation

Abstract

The present disclosure relates to methods for producing engineered eukaryotic cells by inducing modifications at target endogenous nucleic acid sequences. Methods disclosed herein may comprise introducing a transfection mixture into a eukaryotic cell via electroporation. In some embodiments, the transfection mixture may comprise a Cas9/sgRNA complex and a cell-free buffer. The Cas9/sgRNA complex may comprise a Cas9 protein and a sgRNA, wherein the sgRNA comprises a crRNA and a tracrRNA. Upon introduction into the eukaryotic cell, the Cas9/sgRNA complex of the transfection mixture may induce a modification of a target endogenous DNA sequence in the nucleus of the eukaryotic cell to provide for an engineered eukaryotic cell

Dependencies are inferred from the claim text.