US 2026/0308549 A1 · GENETICALLY-MODIFIED T CELLS COMPRISING A MODIFIED INTRON IN THE T CELL RECEPTOR ALPHA GENE

Abstract

The present invention provides a genetically-modified T cell comprising in its genome a modified human T cell receptor alpha gene. The modified T cell receptor alpha gene comprises an exogenous sequence of interest inserted into an intron within the T cell receptor alpha gene that is positioned 5′ upstream of TRAC exon 1. The exogenous sequence of interest can comprise an exogenous splice acceptor site and/or a poly A signal, which disrupts expression of the T cell receptor alpha subunit. The sequence of interest can also include a coding sequence for a polypeptide, such as a chimeric antigen receptor. Additionally, the endogenous splice donor site and the endogenous splice acceptor site flanking the intron are unmodified and/or remain functional. The invention further provides compositions and methods for producing the genetically-modified cell, and populations of the cell, and methods for the treatment of a disease, such as cancer, using such cells.

Claims16

Dependent claims

A method for producing a genetically-modified human T cell comprising a modified human T cell receptor alpha gene, said method comprising:

(a) introducing into a human T cell:
(i) a polynucleotide comprising a first nucleic acid sequence encoding an engineered nuclease, wherein said engineered nuclease is expressed in said human T cell;
or
(ii) an engineered nuclease protein;
and
(b) introducing into said human T cell a second polynucleotide comprising a second nucleic acid sequence comprising an exogenous sequence of interest;
wherein said engineered nuclease produces a cleavage site at a recognition sequence within an intron in said human T cell receptor alpha gene that is positioned 5′ upstream of the T cell receptor alpha constant region exon 1,
and wherein said sequence of interest is inserted into said human T cell receptor alpha gene at said cleavage site,
and wherein said sequence of interest comprises an exogenous splice acceptor site and a poly A signal,
and wherein the endogenous splice donor site and the endogenous splice acceptor site flanking said intron are unmodified.

7 more elements

Dependencies are inferred from the claim text.

Genetically-modified t cells comprising a modified intron in the t cell receptor alpha gene

Life of this patent

Genetically-modified t cells comprising a modified intron in the t cell receptor alpha gene

Life of this patent

Abstract

Claims16