Functional single-cell genomics of human cytomegalovirus infection

Understanding how viral and host factors interact and how perturbations impact infection is the basis for designing antiviral interventions. Here we define the functional contribution of each viral and host factor involved in human cytomegalovirus infection in primary human fibroblasts through pooled CRISPR interference and nuclease screening. To determine how genetic perturbation of critical host and viral factors alters the timing, course and progression of infection, we applied Perturb-seq to record the transcriptomes of tens of thousands of CRISPR-modified single cells and found that, normally, most cells follow a stereotypical transcriptional trajectory. Perturbing critical host factors does not change the stereotypical transcriptional trajectory per se but can stall, delay or accelerate progression along the trajectory, allowing one to pinpoint the stage of infection at which host factors act. Conversely, perturbation of viral factors can create distinct, abortive trajectories. Our results reveal the roles of host and viral factors and provide a roadmap for the dissection of host-pathogen interactions. Pooled and single-cell CRISPR screens provide a detailed map of human cytomegalovirus infection.

Automated summary

Understanding the roles of viral and host factors in human cytomegalovirus infection was explored using CRISPR interference and nuclease screening. The study revealed how perturbations of critical host and viral factors can alter the progression and timing of infection. Results showed that perturbing host factors can impact the stage of infection, while perturbation of viral factors can lead to distinct infection trajectories.