Multiplex genome editing of immune cells to enhance functionality and resistance to suppressive environment
AssigneeBoard of Regents, The University of Texas System
InventorsRafet Basar, Elizabeth Shpall, Katy Rezvani
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Abstract
Provided herein are methods for producing immune cells with disruption of multiple genes. Further provided are methods for inserting a chimeric antigen receptor at a gene locus of an immune cell.
Claims148
An in vitro method for the disruption of at least two genes in an immune cell, wherein the at least two genes are selected from the group consisting of NKG2A, SIGLEC-7, LAG3, TIM3, CISH, FOXO1, TGFBR2, TIGIT, CD96, ADORA2, NR3C1, PD1, PDL-1, PDL-2, CD47, SIRPA, SHIP1, ADAM17, RPS6, 4EBP1, CD25, CD40, IL21R, ICAM1, CD95, CD80, CD86, IL10R, CD5, and CD7.
An immune cell with disrupted expression of at least two genes in the immune cell, wherein the at least two genes are selected from the group consisting of NKG2A, SIGLEC-7, LAG3, TIM3, CISH, FOXO1, TGFBR2, TIGIT, CD96, ADORA2, NR3C1, PD1, PDL-1, PDL-2, CD47, SIRPA, SHIP1, ADAM17, RPS6, 4EBP1, CD25, CD40, IL21R, ICAM1, CD95, CD80, CD86, IL10R, CD5, and CD7.
The cell of any one of claims 83-90, wherein the CAR comprises at least one signaling domain selected from the group consisting of CD3, CD28, OX40/CD134, 4-1BB/CD137, FcεRIγ, ICOS/CD278, ILRB/CD122, IL-2RG/CD132, DAP12, CD70, and CD40.
An expression vector encoding a CAR, inhibitory gene sequence, and gRNA.
A method for engineering an immune cell to express a CAR, said method comprising using a CRISPR gRNA to insert the CAR at an inhibitory gene locus of said immune cell.
An immune cell with a CAR inserted at an inhibitory gene of said immune cell.
Dependencies are inferred from the claim text.