Patent ID: 8609419
Filing Date: 2013-12-17
Classification: C12N

Abstract:
1. A method for in vitro insertion of a nucleic acid of interest initially included in a DNA vector, within a predetermined target nucleotide sequence present in a chromosome contained in a target prokaryotic or eukaryotic cell genome that does not comprise said nucleic acid of interest at said target nucleotide sequence, said method comprising: a) providing a DNA vector that comprises the nucleic acid of interest, the DNA vector being replication competent when in said target prokaryotic or eukaryotic cell, b) contacting said DNA vector with a mutagenic agent blocking intracellular DNA replication of said DNA vector to produce a modified DNA vector; c) transfecting said target prokaryotic or eukaryotic cells with the modified DNA vector obtained at the end of step b) under conditions wherein replication of said modified DNA vector commences and insertion of the nucleic acid of interest within said predetermined target nucleotide sequence occurs; and d) selecting prokaryotic or eukaryotic cells for which the nucleic acid of interest has been integrated into the predetermined target nucleotide sequence as a result of said transfecting of step c).