US 12,234,461 B2
Targeted non-viral DNA insertions
Theodore Lee Roth, San Francisco, CA (US); and Alexander Marson, San Francisco, CA (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, A CALIFORNIA CORPORATION
Filed by The Regents of the University of California, Oakland, CA (US)
Filed on Sep. 8, 2023, as Appl. No. 18/463,589.
Application 18/463,589 is a division of application No. 17/498,531, filed on Oct. 11, 2021, granted, now 11,814,624.
Application 17/498,531 is a continuation of application No. 16/622,843, abandoned, previously published as PCT/US2018/037919, filed on Jun. 15, 2018.
Claims priority of provisional application 62/552,180, filed on Aug. 30, 2017.
Claims priority of provisional application 62/520,117, filed on Jun. 15, 2017.
Prior Publication US 2024/0117360 A1, Apr. 11, 2024
Int. Cl. C12N 15/113 (2010.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/1138 (2013.01) [C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/90 (2013.01); C12N 2310/20 (2017.05); C12N 2320/53 (2013.01)] 20 Claims
 
1. An isolated viable, primary human immune cell comprising a complex comprising a ribonucleoprotein complex (RNP) and a DNA template, wherein the RNP comprises a nuclease domain and a guide RNA, wherein the size of the DNA template is greater than or equal to 300 base pairs (bp) in size, and wherein the 5′ and 3′ ends of the DNA template comprise nucleotide sequences that are homologous to
genomic sequences flanking an insertion site in the genome of the primary cell, and wherein the primary cell does not comprise a viral vector for delivery of the DNA template.