STING gain-of-function disrupts lymph node organogenesis and innate lymphoid cell differentiation in mice. Bennion et al
Description
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.