US 12,442,016 B2
Scarless genome editing through two-step homology directed repair
Kazuya Ikeda, Koto (JP); and Matthew H. Porteus, Stanford, CA (US)
Assigned to The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
Filed by The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
Filed on May 18, 2023, as Appl. No. 18/320,132.
Application 18/320,132 is a continuation of application No. 16/740,234, filed on Jan. 10, 2020, granted, now 11,692,202.
Application 16/740,234 is a continuation of application No. PCT/US2018/042702, filed on Jul. 18, 2018.
Claims priority of provisional application 62/533,780, filed on Jul. 18, 2017.
Prior Publication US 2023/0348932 A1, Nov. 2, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C07H 21/02 (2006.01); C07H 21/04 (2006.01); C12N 9/22 (2006.01); C12N 15/85 (2006.01); C12N 15/90 (2006.01); G01N 21/64 (2006.01); A61K 48/00 (2006.01); C12N 5/0735 (2010.01); C12N 5/074 (2010.01)
CPC C12N 15/85 (2013.01) [C12N 9/22 (2013.01); C12N 15/902 (2013.01); C12N 15/907 (2013.01); G01N 21/6486 (2013.01); A61K 48/00 (2013.01); C12N 5/0606 (2013.01); C12N 5/0607 (2013.01); C12N 2310/122 (2013.01); C12N 2310/531 (2013.01); C12N 2510/04 (2013.01)] 14 Claims
 
1. A method for selection marker removal from a genome of a cell comprising performing a combination of a double-stranded break at a target sequence in the genome of the cell, homology directed repair-driven removal of at least one selection marker at the target sequence, and negative selection for the at least one selection marker, wherein the double-stranded break is a non-gene destructive double-stranded break.