Altering gene expression in modified T cells and uses thereof
AssigneeThe Trustees of the University of Pennsylvania
InventorsYangbing Zhao, Jiangtao Ren, Xiaojun Liu, Carl H. June
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Abstract
The present invention relates to compositions and methods for generating a modified T cell with a nucleic acid capable of downregulating endogenous gene expression selected from the group consisting of TCR α chain, TCR β chain, beta-2 microglobulin and FAS further comprising a nucleic acid encoding a modified T cell receptor (TCR) comprising affinity for a surface antigen on a target cell or an electroporated nucleic acid encoding a chimeric antigen receptor (CAR). Also included are methods and pharmaceutical compositions comprising the modified T cell for adoptive therapy and treating a condition, such as an autoimmune disease.
Claims16
A method for generating a CRISPR-modified T cell, the method comprising:
- introducing into a T cell a CRISPR system comprising a nucleic acid that causes downregulation of gene expression of an endogenous TCR α chain in the T cell, wherein the nucleic acid that causes downregulation of gene expression of the endogenous TCR α chain comprises a nucleotide sequence encoded by SEQ ID NO:
- 1;
- introducing into the T cell a CRISPR system comprising a nucleic acid that causes downregulation of gene expression of an endogenous TCR β chain in the T cell;
- and
- introducing into the T cell a nucleic acid encoding an 8F T cell receptor (TCR) comprising affinity for NY-ESO-1 on a target cell,
- thereby generating the CRISPR-modified T cell.
A method for generating a CRISPR-modified T cell, the method comprising:
- introducing into a T cell a CRISPR system comprising a nucleic acid that causes downregulation of gene expression of an endogenous TCR α chain in the T cell, wherein the nucleic acid that causes downregulation of gene expression of the endogenous TCR α chain comprises a nucleotide sequence encoded by SEQ ID NO:
- 1;
- introducing into the T cell a CRISPR system comprising a nucleic acid that causes downregulation of gene expression of an endogenous TCR β chain in the T cell;
- introducing into the T cell a CRISPR system comprising a nucleic acid that causes downregulation of gene expression of an endogenous PD1 gene in the T cell;
- and
- introducing into the T cell a nucleic acid encoding an 8F T cell receptor (TCR) comprising affinity for NY-ESO-1 on a target cell,
- thereby generating the CRISPR-modified T cell.
A method for generating a CRISPR-modified T cell, the method comprising introducing into a T cell:
- i) a first nucleic acid encoding Cas9;
- ii) a second nucleic acid comprising a nucleotide sequence encoded by SEQ ID NO:
- 1;
- iii) a third nucleic acid;
- and
- iv) a fourth nucleic acid encoding an 8F T cell receptor (TCR), the TCR comprising affinity for NY-ESO-1;
- wherein, in the introducing:
- i) Cas9 is expressed;
- ii) a nucleotide sequence of the second nucleic acid binds to the Cas9 and the nucleotide sequence of SEQ ID NO:
- 1 hybridizes with an endogenous TCR α chain gene,
- thereby causing downregulation of the endogenous TCR α gene;
- and
- iii) a nucleotide sequence of the third nucleic acid binds to the Cas9 and hybridizes with an endogenous TCR β chain gene, thereby causing downregulation of the endogenous TCR β gene;
- thereby generating the CRISPR-modified T cell.
Dependencies are inferred from the claim text.