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hammerlab / tcr-transfer

Explore an alternative way to clone TILs by first single-cell TCR-seq'ing them and then transforming non-exhausted T cells with these TIL TCRs
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Get the pmel TCR synthesized #13

Closed armish closed 4 years ago

armish commented 4 years ago

Similar to what we have done with the OT-I for @pinarden's paper, we need this TCR in house for better positive/negative control against LLC (see #3).

>pmel_TCRA
ATGATGAAATCCTTGAGTGTTTCACTAGTGGTCCTGTGGCTCCAGTTTAATTGGGTGAGAAGCCAGCAGAAGGTGCAGCAGAGCCCAGAATCCCTCACTGTCTCAGAGGGAGCCATGGCCTCTCTCAACTGCACTTTCAGTGATCGTTCTTCTGACAACTTCAGGTGGTACAGACAGCATTCTGGGAAAGGCCTTGAGGTGCTGGTGTCCATCTTCTCTGATGGTGAAAAGGAAGAAGGCAGTTTTACAGCTCACCTCAATAGAGCCAGCCTGCATGTTTTCCTACACATCAGAGAGCCGCAACCCAGTGACTCTGCTCTCTACCTCTGTGCAGTGAACACAGGAAACTACAAATACGTCTTTGGAGCAGGTACCAGACTGAAGGTTATAGCACACATCCAGAACCCAGAACCTGCTGTGTACCAGTTAAAAGATCCTCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAATCAATGTGCCGAAAACCATGGAATCTGGAACGTTCATCACTGACAAAACTGTGCTGGACATGAAAGCTATGGATTCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTGCCAAGATATCTTCAAAGAGACCAACGCCACCTACCCCAGTTCAGACGTTCCCTGTGATGCCACGTTGACTGAGAAAAGCTTTGAAACAGATATGAACCTAAACTTTCAAAACCTGTCAGTTATGGGACTCCGAATCCTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGATAA 

>pmel_TCRB
ATGGGCACCAGGCTTCTTGGCTGGGCAGTGGGCCCGCGGCTTCTTGGCTGGGCAGTGTTCTGTCTCCTTGACACAGTACTGTCTGAAGCTGGAGTCACCCAGTCTCCCAGATATGCAGTCCTACAGGAAGGGCAAGCTGTTTCCTTTTGGTGTGACCCTATTTCTGGACATGATACCCTTTACTGGTATCAGCAGCCCAGAGACCAGGGGCCCCAGCTTCTAGTTTACTTTCGGGATGAGGCTGTTATAGATAATTCACAGTTGCCCTCGGATCGATTTTCTGCTGTGAGGCCTAAAGGAACTAACTCCACTCTCAAGATCCAGTCTGCAAAGCAGGGCGACACAGCCACCTATCTCTGTGCCAGCAGTTTCCACAGGGACTATAATTCGCCCCTCTACTTTGCGGCAGGCACCCGGCTCACTGTGACAGAGGATCTGAGAAATGTGACTCCACCCAAGGTCTCCTTGTTTGAGCCATCAAAAGCAGAGATTGCAAACAAACAAAAGGCTACCCTCGTGTGCTTGGCCAGGGGCTTCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTCCACAGTGGGGTCAGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAGCAGCCGCCTGAGGGTCTCTGCTACCTTCTGGCACAATCCTCGCAACCACTTCCGCTGCCAAGTGCAGTTCCATGGGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACACAGAACATCAGTGCAGAGGCCTGGGGCCGAGCAGACTGTGGGATTACCTCAGCATCCTATCAACAAGGGGTCTTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAAGCCACCCTGTATGCTGTGCTTGTCAGTACACTGGTGGTGATGGCTATGGTCAAAAGAAAGAATTCATGATAA

Going to flank it with AflII and NotI similar to other TCRs for uniformity.

armish commented 4 years ago

Original sequences:

elinorgottschalk commented 4 years ago

The pmel TCR DNA plasmids arrived on 1/24/2020.