Patent ID: 8685737

Claim:
A method for simultaneously integrating a plurality of (n) exogenous nucleic acids into a plurality of (n) target sites of a host cell genome, wherein n is at least two, the method comprising: (a) simultaneously contacting a host cell with: (i) said plurality of exogenous nucleic acids, wherein: x is an integer that varies from 1 to n, and for each integer x, each exogenous nucleic acid (ES) x comprises a first homology region (HR1) x and a second homology region (HR2) x , wherein (HR1) x and (HR2) x are capable of initiating host cell mediated homologous recombination of (ES) x at a target site (TS) x selected from said plurality of (n) target sites of said host cell genome; and (ii) for each said target site (TS) x , a nuclease (N) x capable of cleaving at (TS) x whereupon said cleaving results in homologous recombination of (ES) x at (TS) x ; and (b) recovering a host cell wherein each exogenous nucleic acid (ES) x has integrated at its selected target site (TS) x , wherein said recovering occurs at a frequency of about one every 10, 9, 8, 7, 6, 5, 4, 3, or 2 contacted host cells, or clonal populations thereof, screened.