Genetics Core
Our Aim:
The aim of this core is to provide a comprehensive and technically advanced resource for the design, generation and production of lentiviralbased shRNA vectors as well as retroviral approaches to manipulate gene expression in primary cells and cell lines of interest. The services provided by this core will include a number of lentiviral vectors and maps, as well as assistance in the identification of candidate shRNA targeting sequences, cloning into viral vectors and validation in the relevant cellular context, by sequencing and assessment of the efficacy at the protein level. This core will also provide training on the proper compliance with biosafety issues required for lentivirus-based research. We will focus our efforts on generating shRNA encoding lentiviruses aimed at knocking down expression of genes that are of specific importance to the participants of this program.shRNA Validation Protocol:
- Search target gene for predicted optimal shRNA sequences.
- Clone primers into shRNA expression plasmid
- Validate knock down of target gene in DO.11.10 T cells
| Core Status as of 5.13.05 | ||
| Gene | Relevant | Status |
| Bim | Apoptosis/Survival | Validated |
| Puma | Apoptosis/Survival | Validated |
| Bax | Apoptosis/Survival | Cloned |
| Bak | Apoptosis/Survival | Cloned |
| Bcl-2 | Apoptosis/Survival | Analyzed |
| Lyn | BCR Signaling | Validated |
| Syk | BCR/TCR Signaling | Validated |
| CD22 | BCR Signaling | Analyzed |
| CD72 | BCR Signaling | Analyzed |
| PKCg | BCR/TCR Signaling | Cloned |
| PKCd | BCR/TCR Signaling | Cloned |
| SHIP | BCR/TCR Signaling | Analyzed |
| SHP1 | BCR/TCR Signaling | Analyzed |
| AKT-1 | Apoptosis/Survival | Analyzed |
| AKT-2 | Apoptosis/Survival | Analyzed |
| AKT-3 | Apoptosis/Survival | Analyzed |
| PTEN | Apoptosis/Survival | Analyzed |
| PDK-1 | Apoptosis/Survival | Analyzed |
| PDK-2 | Apoptosis/Survival | Analyzed |
| mTOR/FRAP | Apoptosis/Survival | Analyzed |
| Foxo-1 | Apoptosis/Survival | Analyzed |
| Foxo-3 | Apoptosis/Survival | Analyzed |
| Foxp-1 | Apoptosis/Survival | Analyzed |
| Foxp-4 | Apoptosis/Survival | Analyzed |
| ZAP70 | TCR Signaling | Analyzed |
| LAT | TCR Signaling | Validated |
| CD8a | TCR Signaling | Validated |
| c-rel | NF-kB Signaling | Validated |
| Rel-a | NF-kB Signaling | Validated |
| Rel-b | NF-kB Signaling | Cloned |
| iKK-Bi | NF-kB Signaling | Cloned |
| IkBe | NF-kB Signaling | Validated |
| IkBa | NF-kB Signaling | Validated |
| Bcl-10 | NF-kB Signaling | Validated |
| IKKa | NF-kB Signaling | Validated |
| IKKb | NF-kB Signaling | Validated |
| NF-kB1 | NF-kB Signaling | Cloned |
| NF-kB2 | NF-kB Signaling | Cloned |
| Bcl-3 | NF-kB Signaling | Cloned |
| Calcineurin | TCR/BCR Signaling | Cloned |
| NFATc1 | TCR Signaling | Cloned |
| NFATc2 | TCR Signaling | Cloned |
| NFATc3 | TCR Signaling | Cloned |
| NFATc4 | TCR Signaling | Cloned |
| Csp1 | TCR/BCR Signaling | Cloned |
| Csp2 | TCR/BCR Signaling | Cloned |
| Csp3 | TCR/BCR Signaling | Cloned |
| Carma | TCR/BCR Signaling | Cloned |
| Calm | TCR/BCR Signaling | Cloned |
| Cabin | TCR/BCR Signaling | Cloned |