| Products/Services Used | Details | Operation |
|---|---|---|
| Custom Vector Construction | Cas9 expressing B cells with stable C-terminal HA epitope-tagged MYC cDNA expression were established by lentiviral transduction and hygromycin selection. MYC cDNA expression was confirmed by immunoblot and by FACS. MYC rescue cDNA, with silent PAM site mutations at MYC sg1 and sg2 sites, is described in the following table. MYC sg1 and sg2 targeting sequences are highlighted in bold. PAM sequences are underlined. Mutation sites are indicated in italic bold. Rescue cDNA was synthesized by GenScript (Piscataway, NJ) and cloned into pLX-TRC313 vector. | Get A Quote |
Epstein-Barr virus (EBV) is associated with multiple human malignancies. To evade immune detection, EBV switches between latent and lyticprograms. How viral latency is maintained in tumors or in memory B cells, the reservoir for lifelong EBV infection, remains incompletely understood. To gain insights, we performed a human genome-wide CRISPR/Cas9 screen in Burkitt lymphoma B cells. Our analyses identified a network of host factors that repress lytic reactivation, centered on the transcription factor MYC, including cohesins, FACT, STAGA, and Mediator. Depletion of MYC or factors important for MYC expression reactivated the lytic cycle, including in Burkitt xenografts. MYC bound the EBV genome orig... More
