Patent ID: 8420395
Filing Date: 2013-04-16
Classification: A61K,C07K,C12N,Y02A

Abstract:
1. A method of site-specifically integrating a polynucleotide sequence of interest into the genome of an isolated cell, said method comprising: introducing into the isolated cell (i) a circular targeting construct comprising a bacterial genomic recombination site (attB) and the polynucleotide sequence of interest, and (ii) a φC31 integrase or an expression cassette encoding the same, wherein the φC31 integrase comprises the amino acid sequence of SEQ ID NO:132, wherein the genome of the isolated eucaryotic cell comprises a pseudo-phage genomic recombination site (pseudo-attP) comprising a nucleic acid sequence of SEQ ID NO:102 and is native to the genome of the cell; and maintaining the isolated cell under conditions that allow recombination between the attB and the pseudo attP, wherein the recombination is mediated by the φC31 integrase and the recombination results in site-specific integration of the polynucleotide sequence of interest in the genome of the isolated cell.