US 12,442,021 B2
Genomic safe harbors for genetic therapies in human stem cells and engineered nanoparticles to provide targeted genetic therapies
Jennifer E. Adair, Seattle, WA (US); and Reza Shahbazi, Seattle, WA (US)
Assigned to Fred Hutchinson Cancer Center, Seattle, WA (US)
Filed by Fred Hutchinson Cancer Center, Seattle, WA (US)
Filed on Dec. 8, 2022, as Appl. No. 18/063,511.
Application 18/063,511 is a continuation of application No. 16/619,211, granted, now 11,603,544, previously published as PCT/US2018/036154, filed on Jun. 5, 2018.
Claims priority of provisional application 62/664,045, filed on Apr. 27, 2018.
Claims priority of provisional application 62/564,129, filed on Sep. 27, 2017.
Claims priority of provisional application 62/515,474, filed on Jun. 5, 2017.
Prior Publication US 2023/0279441 A1, Sep. 7, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C07H 21/02 (2006.01); A61K 40/10 (2025.01); A61K 40/40 (2025.01); A61K 47/69 (2017.01); C07H 21/04 (2006.01); C12N 5/078 (2010.01); C12N 9/22 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01)
CPC C12N 15/907 (2013.01) [A61K 40/10 (2025.01); A61K 40/40 (2025.01); A61K 47/6929 (2017.08); C12N 5/0634 (2013.01); C12N 9/22 (2013.01); C12N 15/113 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); C12N 2510/00 (2013.01)] 20 Claims
 
1. A therapeutic formulation comprising a nanoparticle (NP) and a pharmaceutically acceptable carrier, the NP comprising:
a metallic core associated with a first NP layer and a second NP layer, wherein the first NP layer comprises:
a crRNA with a 3′ end and a 5′ end,
wherein the 3′ end is conjugated to a spacer with a thiol modification, and the 5′ end is conjugated to a nuclease to form a crRNA-nuclease ribonucleoprotein (RNP) complex,
wherein the thiol modification is covalently linked to the surface of the metallic NP core and
wherein the crRNA-nuclease RNP complex binds a target PAM-site within human (h) chromosome (chr) 11 at positions 67681215-67741765; 67691162-67691186; 67723825-67723849; 67805337-67845629; 67812349-67812375; 67812443-67812469; 67839126-67839150; or 67895738-67941098 or hchr3 at positions 46373915-46373939,
and wherein the second NP layer comprises a donor template comprising a therapeutic gene and homology-directed repair templates, and wherein the second NP layer is farther from the surface of the metallic core than the first NP layer.