	US 11,959,093 B2
	Integration sites and uses thereof
Ling-Jie Kong, Union City, CA (US); Ruby Yanru Tsai, San Jose, CA (US); and Xiuling Chi, San Jose, CA (US)
Assigned to APPLIED STEMCELL, INC., Milpitas, CA (US)
Appl. No. 16/492,950
Filed by APPLIED STEMCELL, INC., Milpitas, CA (US)
PCT Filed Mar. 14, 2018, PCT No. PCT/US2018/022274 § 371(c)(1), (2) Date Sep. 11, 2019, PCT Pub. No. WO2018/175153, PCT Pub. Date Sep. 27, 2018.
Claims priority of provisional application 62/473,454, filed on Mar. 19, 2017.
Prior Publication US 2020/0017884 A1, Jan. 16, 2020
Int. Cl. C12N 15/79 (2006.01); C07H 21/04 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01); C07H 21/02 (2006.01); C12N 15/63 (2006.01); C12N 15/85 (2006.01); C12N 15/86 (2006.01)

CPC C12N 15/907 (2013.01) [C12N 15/113 (2013.01); C07H 21/02 (2013.01); C07H 21/04 (2013.01); C12N 15/63 (2013.01); C12N 15/79 (2013.01); C12N 15/85 (2013.01); C12N 15/86 (2013.01); C12N 2310/20 (2017.05)]

10 Claims

1. A method, comprising:

introducing an exogenous nucleic acid into an isolated mammalian cell in vitro; and

integrating the exogenous nucleic acid into a locus of the genome of the isolated mammalian cell using

(i) a Cas nuclease and a gRNA targeting the locus of the genome, or

(ii) a recombinase,

wherein the locus comprising an extended methylation-free CpG island and a nucleotide sequence at least 90% identical to SEQ ID NO:1.