STING gain-of-function disrupts lymph node organogenesis and innate lymphoid cell differentiation in mice. Bennion et al
Heterozygous STING N153S mutant mice lack lymph nodes and Peyer’s patches. Lymph node development requires a RORT+ lymphoid tissue inducer (LTi) cells. In STING N153S fetuses, there are fewer LTi cells. These LTi cells arise in the fetal liver from CD127+ 47+ progenitor cells. We hypothesized that STING N153S impairs LTi cell survival or function, and that transcriptional abnormalities might be detectable by single-cell RNA-sequencing. To enrich for this rare cell type, single Lin–CD45+cKITintCD127+47+ cells were sorted directly into a 96-well plate (one cell per well) containing 2 l of 10x lysis buffer (Takara catalog no. 635013) and 5% RNAse inhibitor (Promega catalog no. PRN2611). After sorting, plates were immediately frozen at -80 degrees Celsius. Single-cell RNA sequencing of each well was performed at the Genome Technology Access Center (GTAC) at Washington University in St. Louis. A total of 48 WT and 48 STING N153S cells were sequenced.