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| Status |
Public on Oct 03, 2022 |
| Title |
speedingCARs: accelerating the engineering of CAR T cells by signaling domain shuffling and single-cell sequencing |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Other
|
| Summary |
Due to the low-throughput process of systematically designing and functionally testing chimeric antigen receptors (CARs), only a small set of immune signaling domains have been thoroughly explored, despite their major role in T cell activation, effector function and persistence. SpeedingCARs presents an integrated method for engineering CAR T cells by signaling domain shuffling and functional screening by single-cell sequencing. Leveraging the inherent modularity of natural signaling domains, we generated a diverse library of 180 unique CAR variants, which were genomically integrated into primary human T cells by CRISPR-Cas9. Functional and pooled screening of the CAR T cell library was performed by co-culture with tumor cells, followed by single-cell RNA sequencing (scRNA-seq) and single-cell CAR sequencing (scCAR-seq), thus enabling high-throughput profiling of multi-dimensional cellular responses.
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| Overall design |
A library of 180 unique CAR variants was genomically integrated into primary human T cells by CRISPR-Cas9 genome editing. Single-cell RNA sequencing data of pooled library CAR T cells from 3 different healthy donors was generated following 36h of co-culture with HER2 expressing breast cancer cell line SKBR3. As negative controls TCR negative T cells and CD28-CD3Z unstimulated CAR T cells were also sequenced.
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| Contributor(s) |
Castellanos-Rueda R, Di Roberto RB, Reddy ST |
| Citation(s) |
36323661 |
| |
| Submission date |
Sep 26, 2022 |
| Last update date |
Nov 17, 2022 |
| Contact name |
Rocio Castellanos Rueda |
| E-mail(s) |
rocio.castellanos@bsse.ethz.ch
|
| Organization name |
ETH
|
| Department |
D-BSSE
|
| Lab |
Reddy LAB
|
| Street address |
Mattenstrasse 26
|
| City |
Basel |
| State/province |
--None-- |
| ZIP/Postal code |
4058 |
| Country |
Switzerland |
| |
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| Platforms (3) |
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| Samples (14)
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| Relations |
| BioProject |
PRJNA884406 |
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