pAAV_uTCR-KI
(Plasmid #257993)

• Purpose: AAV transfer plasmid cloning backbone for TCR knock-in mouse generation.
• Depositing Lab: Gabriel Victora
• Publication: Bilanovic et al EMBO J. 2026 May 13. doi: 10.1038/s44318-026-00779-z.

Backbone

• Vector backbone: pAAV-RC6
• Backbone manufacturer: Cell biolabs
• Modifications to backbone: S663V + T492V (Pandya M, Britt K, Hoffman B, Ling C, Aslanidi GV (2015) Reprogramming immune response with capsid-optimized AAV6 vectors for immunotherapy of cancer. J Immunother 38:292–298) F129L + Y445F + Y731F (van Lieshout LP, Domm JM, Rindler TN, Frost KL, Sorensen DL, Medina SJ, Booth SA, Bridges JP, Wootton SK (2018) A novel triple-mutant AAV6 capsid induces rapid and potent transgene expression in the muscle and respiratory tract of mice. Mol Ther Methods Clin Dev 9:323–329) Y705 + Y731F + T492V (Ling C, Bhukhai K, Yin Z, Tan M, Yoder MC, Leboulch P, Payen E, Srivastava A (2016) High-efficiency transduction of primary human hematopoietic stem/ progenitor cells by AAV6 vectors: strategies for overcoming donor-variation and implications in genome editing. Sci Rep 6:35495)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Trbv16 promoter flanked by 5' and 3' homology arms to the endogenous Trbj2 locus
• Species: M. musculus (mouse)
• Insert Size (bp): 2050

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The pAAV-RC6 (cat. no. VPK-426) plasmid was purchased from Cell Biolabs and we modified it as indicated.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

For cloning, digest the plasmid with EcoRI and insert desired TCRα VJ-C and TCRβ VDJ sequences, separated by a P2A ribosome-skipping peptide using gibson assembly. When designing the insert, add the designated 45bps "homology cloning insert" the 5' and 3' of your gene fragments to ensure proper assembly.