T cell receptors and methods of use thereof

A nucleic acid molecule comprising (i) a first nucleotide sequence encoding a recombinant T cell receptor (TCR) or an antigen binding portion thereof that specifically binds human gp100 (“anti-gp100 TCR”);

• and (ii) a second nucleotide sequence, wherein the second nucleotide sequence or the polypeptide encoded by the second nucleotide sequence inhibits the expression of an endogenous TCR,
• wherein the anti-gp100 TCR binds the same epitope or an overlapping epitope of human gp100 as a reference TCR, which comprises an alpha chain and a beta chain, wherein the alpha chain comprises an amino acid sequence as set forth in SEQ ID NO:
• 1 and the beta chain comprises an amino acid sequence as set forth in SEQ ID NO:
• 2.

A recombinant T cell receptor (TCR) or an antigen binding portion thereof that specifically binds human gp100 (“an anti-gp100 TCR”), which cross competes for binding to human gp100 with a reference TCR;

• wherein the reference TCR comprises an alpha chain and a beta chain, and wherein the alpha chain comprises an amino acid sequence as set forth in SEQ ID NO:
• 1 and the beta chain comprises an amino acid sequence as set forth in SEQ ID NO:
• 2;
• and
• wherein the anti-gp100 TCR comprises an alpha chain and a beta chain, wherein the alpha chain comprises a constant region, and wherein the beta chain comprises a constant region;
• wherein
• (i) the alpha chain constant region comprises an amino acid sequence having a least 1, at least 2, at least 3, at least 4, or at least 5 amino acid substitutions relative to a constant region present in the amino acid sequence set forth in SEQ ID NO:
• 1 or
• (ii) the beta chain constant region comprises an amino acid sequence having a least 1, at least 2, at least 3, at least 4, or at least 5 amino acid substitutions relative to a constant region present in the amino acid sequence of SEQ ID NO:
• 2.

A recombinant T cell receptor (TCR) or an antigen binding portion thereof that specifically binds human gp100 (“an anti-gp100 TCR”), which binds the same epitope or an overlapping epitope of human gp100 as a reference TCR;

• wherein the reference TCR comprises an alpha chain and a beta chain, and wherein the alpha chain comprises an amino acid sequence as set forth in SEQ ID NO:
• 1 and the beta chain comprises an amino acid sequence as set forth in SEQ ID NO:
• 2;
• an
• wherein the anti-gp100 TCR comprises an alpha chain and a beta chain, wherein the alpha chain comprises a constant region, and wherein the beta chain comprises a constant region;
• wherein
• (i) the alpha chain constant region comprises an amino acid sequence having a least 1, at least 2, at least 3, at least 4, or at least 5 amino acid substitutions relative to a constant region present in the amino acid sequence set forth in SEQ ID NO:
• 1 or
• (ii) the beta chain constant region comprises an amino acid sequence having a least 1, at least 2, at least 3, at least 4, or at least 5 amino acid substitutions relative to a constant region present in the amino acid sequence set forth in SEQ ID NO:
• 2.