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MB Sample ID: SA343040
Local Sample ID: | D16 19BBz_2 |
Subject ID: | SU003286 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
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Treatment:
Treatment ID: | TR003295 |
Treatment Summary: | The human CD19-BBz shRNA CAR lentiviral constructs were based on a previously described CD19-BBz CAR containing the human CD19-binding scFV FMC63, CD8 hinge domain, 4-1BB costimulatory domain and CD3 chain105. Using CD19-BBz CAR plasmid as a cloning vector, we generated multiple vectors where we incorporated the RNA polymerase III U6 promotor (on the 3' of the CD3 chain domain) followed by an shRNA: 1) a CD19-BBz/shMCJ-1 CAR construct containing the shMCJ-1 5’-GAAGATTTCAACTCCTAGC-3’ sequence106, 2) a CD19-BBz/shMCJ-2 CAR construct containing the shMCJ-2; 5’-AACCTCTAGAACAAGTTATC-3’, and 3) a CD19-BBz/c-shRNA CAR vectors expressing the shRNA encoding scramble sequences. Lentiviral supernatant was produced in the LentiX-293T packaging cell line (Clonetech) as previously described. Lentiviral supernatants were collected after 48 hours post-transfection. |