Genomic safe harbors for genetic therapies in human stem cells and engineered nanoparticles to provide targeted genetic therapies
AssigneeFred Hutchinson Cancer Research Center
InventorsJennifer E. Adair, Reza Shahbazi
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Abstract
Genomic safe harbors (GSH) for genetic therapies in human stem cells and engineered nanoparticles to provide targeted genetic therapies are described. The GSH and/or associated nanoparticles can be used to safely and efficiently treat a variety of genetic, infectious, and malignant diseases.
Claims41
A method of genetically modifying a blood cell through homology-directed repair (HDR) within a universal hematopoietic stem cell (HSC) safe harbor loci of SEQ ID NO:
- 132 wherein the method comprises:
- contacting the blood cell with a gold nanoparticle (AuNP) associated with at least two active layers wherein
- the first 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 Cpf1, and wherein the thiol modification is covalently linked to the surface of the AuNP and wherein the crRNA comprises SEQ ID NO:
- 195;
- and
- the second layer comprises a donor template comprising a therapeutic gene and homology-directed repair templates;
- and wherein the second layer is farther from the surface of the AuNP than the first layer
- thereby genetically modifying the blood cell through HDR within a universal HSC safe harbor loci of SEQ ID NO:
- 132.
A gold nanoparticle (AuNP) associated with two active layers wherein
- the first 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 Cpf1, and wherein the thiol modification is covalently linked to the surface of the AuNP and wherein the crRNA comprises SEQ ID NO:
- 195;
- and
- the second layer comprises a donor template comprising a therapeutic gene and homology-directed repair templates;
- and wherein the second layer is farther from the surface of the AuNP than the first layer.
A method of genetically modifying a blood cell at a target sequence within chromosome 11 wherein the target sequence comprises SEQ ID NOs. 1-194, 197-208, 210, 213, 242, 245, 251, or 254, comprising:
- contacting the blood cell with a targeting element, a cutting element, and a homology-directed repair template wherein the contacting results in (i) cutting within the target sequence;
- and (ii) homology-directed repair (HDR).
A nanoparticle associated with at least two active layers wherein the first layer comprises a DNA targeting element and a cutting element and wherein the second layer comprises a donor template comprising a therapeutic gene and homology-directed repair templates;
- and wherein at least portions of the second layer are farther from the surface of the nanoparticle than the first layer and wherein the targeting element hybridizes to one of SEQ ID NOs. 1-194, 197-208, 210, 213, 242, 245, 251, 254, 258, or 263.
Dependencies are inferred from the claim text.