Versatile and efficient non-viral integration of large transgenes in human T cells via CRISPR knock-in and engineered integrases
Kassing I, Kath J, Nitulescu A-M, Glaser V, Hartmann LM, Pu Y, Huth L, Kārkliņš R, Shaji S, Ringel AR, Pouzolles M, Stein M, Ibrahim DM, Wagner DL
bioRxiv 2025.09.10.675267
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Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 247037 | PASTA-1_pUC19_HDR-Bxb1-attG_TRAC_for-Cas12a | Encodes HDR-template for Cas12a-mediated Knock-In of Bxb1-attG (landing pad) into TRAC gene (exon1). Template region to be PCR-amplified. |
| 247038 | PASTA-2_trunc-pUC19_INT-Bxb1-attV_P2A_CD19CAR_bGH-pA | Integration-Plasmid with Bxb1-attV (docking site) for bxb1-mediated integration of a CD19-CAR. Targeted whole plasmid insertion after maxiprep + AMPure bead purification. |
| 247039 | PASTA-3_pUC19_HDR-Pa01-attG_TRAC_for-Cas12a | Encodes HDR-template for Cas12a-mediated Knock-In of size-reduced Pa01-attG (landing pad) into TRAC gene (exon1). Template region to be PCR-amplified. |
| 247040 | PASTA-4_pUC19_INT-Pa01-attV_P2A_CD19CAR_bGH-pA | Integration-Plasmid with Pa01-attV (docking site) for Pa01-mediated integration of a CD19-CAR. Targeted whole plasmid insertion after maxiprep + AMPure bead purification. |
| 247041 | PASTA-5_pUC19_IVT-ePa01 | Plasmid for in vitro transcription (IVT) of enhanced Pa01 (ePa01) mRNA. Use indicated PCR-primers for correction of T7-mutation and addition of poly-A-tail. |