Enhanced gene expression

摘要

The present disclosure is directed to a novel, unexpected approach of expressing exogenous gene(s) at increased levels by predictably, optionally irreversibly, incorporating the gene(s) into a region of increased gene expression (RIDGE) on a chromosome of a host cell. This approach is accomplished by identification of RIDGE(s) and further by integration of integrase-specific sites (e.g., attP or attB) in the presence of or mediated by integrase. The approach renders a high level of gene expression; predictability of the location of the exogenous genes, elimination of genetic instability or unwanted phenotype; and reduction of time and cost in optimizing protein production in host cells, which will be every useful in the production of therapeutic, prophylactic, and diagnostic proteins.

主权项

1. An isolated host cell comprising
an exogenous gene flanked by a first integrase-resulting site and a second integrase-resulting site; and a fraction of green fluorescent protein (GFP) gene sequence, wherein the exogenous gene and the fraction of GFP gene sequence are located at a region of increased gene expression (RIDGE) on a chromosome of the cell, and wherein the fraction of GFP gene sequence does not express GFP.