US 11,795,479 B2
CRISPR enabled multiplexed genome engineering
Andrew Garst, Boulder, CO (US); and Ryan T Gill, Denver, CO (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF COLORADO, Denver, CO (US)
Filed by The Regents of the University of Colorado, Denver, CO (US)
Filed on Apr. 5, 2021, as Appl. No. 17/222,936.
Application 17/222,936 is a continuation of application No. 16/938,739, filed on Jul. 24, 2020, granted, now 11,078,498.
Application 16/938,739 is a continuation of application No. 16/550,092, filed on Aug. 23, 2019.
Application 16/550,092 is a continuation of application No. 16/275,439, filed on Feb. 14, 2019, granted, now 10,435,715, issued on Oct. 8, 2019.
Application 16/275,439 is a continuation of application No. 16/056,310, filed on Aug. 6, 2018, granted, now 10,364,442, issued on Jul. 30, 2019.
Application 16/056,310 is a continuation of application No. 15/948,789, filed on Apr. 9, 2018, granted, now 10,240,167, issued on Jul. 16, 2019.
Application 15/948,789 is a continuation of application No. 15/948,785, filed on Apr. 9, 2018, granted, now 10,351,877, issued on Mar. 26, 2019.
Application 15/948,785 is a continuation of application No. 15/630,909, filed on Jun. 22, 2017, granted, now 9,982,278, issued on May 29, 2018.
Application 15/630,909 is a continuation of application No. 15/116,616, granted, now 10,266,849, issued on Apr. 23, 2019, previously published as PCT/US2015/015476, filed on Feb. 11, 2015.
Claims priority of provisional application 61/938,608, filed on Feb. 11, 2014.
Prior Publication US 2021/0254104 A1, Aug. 19, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/90 (2006.01); C12N 15/10 (2006.01); C40B 40/08 (2006.01); C12N 15/11 (2006.01); C12N 9/22 (2006.01)
CPC C12N 15/902 (2013.01) [C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/1024 (2013.01); C12N 15/1082 (2013.01); C12N 15/1093 (2013.01); C12N 15/11 (2013.01); C12N 15/111 (2013.01); C40B 40/08 (2013.01); C12N 2310/10 (2013.01); C12N 2310/20 (2017.05); C12N 2320/11 (2013.01); C12N 2320/12 (2013.01); C12N 2330/31 (2013.01)] 20 Claims
 
1. A vector for editing a cell created by:
ligating into the vector that comprises a first region of a gRNA a second nucleotide sequence comprising:
a region homologous to a target region in a cell comprising a change in sequence relative to the target region;
a site conferring immunity to nuclease-mediated editing; and
a nucleic acid sequence comprising a second region of the gRNA, whereupon the ligation forms a sequence within the vector comprising a functional gRNA;
thus generating the vector for editing the cell.