Patent ID: 11866726
Assignee: EDITAS MEDICINE, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 8:
9. The method of claim 7, wherein:
(i) the first stuffer has a sequence having less than 50% sequence identity to any nucleic acid sequence within 500 base pairs of the cleavage site, and wherein the second stuffer has a sequence having less than 50% sequence identity to any nucleic acid sequence within 500 base pairs of the cleavage site;
(ii) the first stuffer has a sequence comprising at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-101, and wherein the second stuffer has a sequence comprising at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-101;
(iii) the first stuffer has a sequence that is not the same as the sequence of the second stuffer;
(iv) when the altered nucleic acid comprises a targeted integration genome editing event at the cleavage site, amplifying the altered nucleic acid using the first primer and the second primer produces a first amplicon, wherein the first amplicon has a sequence that is substantially identical to a sequence consisting of either (a) the first donor homology arm and the first stuffer, or (b) the second stuffer and the second donor homology arm;
(v) the cell is a population of cells, and wherein, when the altered nucleic acid in all the cells within the population of cells comprises a targeted integration genome editing event at the cleavage site, amplifying the altered nucleic acid using the first primer and the second primer produces a first amplicon, wherein the first amplicon has a sequence that is substantially identical to a sequence consisting of either i) the first donor homology arm and the first stuffer, or ii) the second stuffer and the second donor homology arm; or
(vi) when the cell is a population of cells, wherein, when the altered nucleic acid in a first cell within the population of cells comprises a non-targeted integration genome editing event at the cleavage site, amplifying the altered nucleic acid using the first primer and the second primer produces a first amplicon, wherein the first amplicon has a sequence that comprises an indel as compared to a sequence of the target nucleic acid; and
wherein, when the altered nucleic acid in a second cell within the population of cells comprises a targeted integration genome editing event at the cleavage site, amplifying the altered nucleic acid in the second cell using the first primer and the second primer produces a second amplicon, wherein the second amplicon has a sequence that is substantially identical to a sequence consisting of either (a) the first donor homology arm and the first stuffer, or (b) the second stuffer and the second donor homology arm.