Poster #050
Optimizing CRISPRi/a Conditions to Study CMV IE2 Latency Circuitry
Mentors: Sonali Chaturvedi, PhD and Yash Chaudhary, PhD
Cytomegalovirus (CMV), infecting more than 70% of the world population, is the leading cause of birth defects and transplant failure. Currently with no cure or vaccine, CMV remains latent in the body and reactivates when the immune system weakens. To study the regulatory mechanisms behind CMV latency, we focused on the virus’s immediate early-2 (IE2) feedback circuit, which plays a critical role in viral fate decision (Chaturvedi et al. Cell 2022). We created isoclonal cell lines for IE2 feedback circuits in human foreskin fibroblast (HFF), retinal epithelial cells (ARPE-19) and human embryonic kidney (HEK 293) cells. Continuing this effort, we are now optimizing conditions for the human genome-wide CRISPR interference and activation (CRISPRi/a) to modulate host gene expression. Notably, this work will lead to a better understanding of how CMV establishes latency with the goal of advancing targeted therapies to improve the safety of stem cell transplantation.