Patent ID: 9340794
Filing Date: 2016-05-17
CPC Classification: C12N

Claim Text:
1. A method of inserting a plurality of nucleic acid molecules in a predetermined order into a predetermined site in the genome of a cell, the method comprising, providing a plurality of nucleic acid molecules to be inserted into a predetermined site in the genome of the cell in a predetermined order next to each other, the plurality of nucleic acid molecules including a first nucleic acid molecule in the order, a last nucleic acid molecule in the order and at least one intervening nucleic acid molecule to be inserted between the first and last nucleic acid molecules, wherein each of the plurality of nucleic acid molecules contains (a) a nucleic acid sequence operatively linked to a promoter sequence at the 5′ end of the nucleic acid molecule, and (b) an overlapping sequence at the 3′ end of the nucleic acid molecule, the promoter sequence in each nucleic acid molecule being different, wherein the overlapping sequence in the at least one intervening nucleic acid molecule is homologous to a portion of a promoter sequence in an adjacent nucleic acid molecule and a portion of the promoter sequence in the at least one intervening nucleic acid molecule is homologous to the overlapping sequence in another adjacent nucleic acid molecule, and wherein a portion of the promoter sequence of the first nucleic acid molecule is homologous to a first sequence in the predetermined site and the overlapping sequence of the last nucleic acid molecule is homologous to a second sequence in the predetermined site; and introducing the plurality of nucleic acid molecules simultaneously into the cell, whereby the plurality of nucleic acid molecules join together in the predetermined order via homologous recombination between the overlapping sequences and the promoter sequences, and are inserted into the genome via homologous recombination between the promoter sequence of the first nucleic acid molecule and the first sequence in the predetermined site and between the overlapping sequence of the last nucleic acid molecule and the second sequence in the predetermined site.