TCR-mRNA construct designTCRshows

Assemble both full-length TCR α and β chains from V/J gene calls and a CDR3, attach the constant regions, join them into a single open reading frame with a 2A peptide, run the checks, and send the result on for codon optimisation and mRNA construct assembly. Aimed at transient TCR-T by mRNA electroporation — nothing integrates into the genome, and the expression window is under your control.

1TCR sequences

V and J gene sequences come from IMGT — the same data the sequence assembler uses, so both pages assemble identically. Overlap between the CDR3 and the V tail or J head is aligned and collapsed automatically.

Alpha chain

Beta chain

Example:

You can also pick a validated TCR from TCRshows-db and arrive here with the fields already filled.

2Constant regions and the chain linker

Murinisation is the other common route to suppressing mispairing with the endogenous human TCR: the introduced chains preferentially pair with each other. The cost is possible murine epitopes and the immunogenicity risk that comes with them. It can be combined with the extra disulfide bond.

This page only gets the protein sequence right. Codon optimisation, UTR choice, secondary structure and mRNA construct assembly all happen in mRNA sequence design — the button above carries the assembled ORF across. They are separate because the failure modes are nothing alike: one wrong residue here gives you a non-functional TCR, whereas over there the question is how well it expresses.
What 2A does, as a matter of record: ribosomal skipping leaves the 2A residues (about 20 aa) on the C terminus of the first chain and a single proline on the N terminus of the second. The second chain’s own leader peptide still works, and that proline is removed along with the signal peptide; the residues on the first chain sit in the cytoplasmic tail. This is inherent to a 2A bicistronic construct, not an assembly error.
Whether to add a Furin site: insert an R-X-[K/R]-R site between the upstream chain and the 2A and Furin will cut after the last R in the Golgi, removing that 2A remnant from the upstream chain. A few basic residues still remain, so it is not truly scarless — and on a TCR those residues sit past the cytoplasmic end of the constant region, where most reports see no clear effect. So it is off by default; turn it on if you want it. All results are for research use only.

Sources: V and J gene and allele sequences come from IMGT, the same source as the sequence assembler; constant region sequences come from UniProtKB/Swiss-Prot — human TRAC P01848, TRBC1 P01850, TRBC2 A0A5B9; mouse TRAC P01849, TRBC1 P01852, TRBC2 P01851. The extra disulfide bond follows Kuball et al. 2007 (Blood 109:2331).