RNA-seq profiling of wild type and Trim33-/- dendritic cell-related progenitors

The development of dendritic cells (DCs) is regulated by complex transcriptional networks. DCs originate from the multipotent progenitors (MPPs) in the bone marrow, which could further give rise to common lymphoid progenitors (CLPs) and common dendritic cell progenitors (CDPs). Whereas CDPs, which could be further divided into CD115+ and CD115- populations, give rise to both conventional (cDC) and plasmacytoid DCs (pDCs), CLP is an addtional source of pDCs. Transcriptome profiling of wild type and Trim33-/- MPPs, CD115- CDPs, and CLPs revealed a critical role of TRIM33 in the development of DCs. Wild type (Trim33fl/fl) or Trim33-/- MPPs (Lin- Sca-1+ CD117+ CD34+ FLT3+), CD115- CDPs (Lin- CD11c- FLT3+ CD117int CD115- CD127-), and CLPs (Lin- CD11c- Sca-1+ CD117+ CD127+ FLT3+) were sorted from 6~8-week-old mice bone marrow and subjected to RNA extraction. Trim33 knockout in vivo relied on Cre recombinase-dependent exicision of Trim33 exon2-4, which would result in frameshift and premature translational stop. Three biological repeats were prepared for individual cell types of each genotype. Transcriptome was profiled by RNA-seq at the Beijing Genomics Institution (BGI) on the BGISEQ-500 platform to generate 50 base-pair single end reads.