Patent ID: 7098031

Claim:
A method for randomly integrating a polynucleotide into a genome of an isolated host, said method comprising: a) introducing into said host cell a vector having no free 5′ and 3′ ends and comprising said polynucleotide, said vector comprising at least one cleavage site for meganuclease, wherein less than 5 copies of said at least one cleavage site are present in the host cell genome; b) causing cleavage of said at least one meganuclease cleavage site in said host cell by said meganuclease, said meganuclease being either introduced into said host cell or expressed by said host cell from a vector comprising a nucleic acid encoding said meganuclease, thereby creating or releasing said polynucleotide in a linear form having free 5′ and 3′ ends from said vector in said host cell; and, c) maintaining the host cell under conditions and for a period of time sufficient to cause the random integration of said created or released polynucleotide into said host cell genome.