Project description:We performed bulk RNA sequencing of several subpopulations of adipose tissue stromal cells to better understand the function of preadipocyte subpopulation (P1 and P2) in mouse inguinal adipose tissue. P1 and P2 cells isolated from inguinal adipose tissue from male and female C57/B6 mice, collected by FACS. P1 cells are CD31-CD45-TER119-SCA1+CD55+VAP1-CD142- cell population, and P2 cells are CD31-CD45-TER119-SCA1+CD55-VAP1+CD142- cell population. 50,000 cells were collected for each populations.
Project description:A human embryonic fibroblast cell line was synchronously infected with poliovirus in the absence or presence of interferon-α, or with vacciniavirus, a virus that is not inhibited by interferon. The titers were sufficient to yield productive infection in a majority of the cells. The cells were harvested in triplicate at various time-points, and the transcriptosome compared with mock infected cells using oligo-based 35 k microarrays. The project had two purposes: to characterize the cellular response and to look for candidate genes involved in viral defense. The changes in gene expression due to vaccinia virus did not correspond to those caused by poliovirus. More surprisingly, neither did the changes when comparing 8 h and 16 h of poliovirus infection. However, a large proportion of the genes up-regulated by interferon-α were also up-regulated by poliovirus, both at 8 h and 16 h. Interferon-α inhibited poliovirus replication, thus the observations suggest that the cells do launch an antiviral response to poliovirus. Moreover, as interferon genes were not induced, the data indicate that several of the relevant genes can be activated in an interferon independent manner. Further analyses of the data led to a list of candidate antiviral genes. Functional information was limited, or absent, for most of these genes. Keywords: Poliovirus; Vacciniavirus; Interferon; Microarray; Gene expression; Defense genes
Project description:Like all biological populations, viral populations exist as networks of genotypes connected through mutation. Mapping the topology of these networks and quantifying population dynamics across them is crucial to understanding how populations adapt to changes in their selective environment. The influence of mutational networks is especially profound in viral populations which rapidly explore their mutational neighborhoods via high mutation rates. Using a novel single-cell sequencing method, scRNAseq-Enabled Acquisition of mRNA and Consensus Haplotypes Linking Individual Genotypes and Host Transcriptomes (SEARCHLIGHT), we captured and assembled viral haplotypes from hundreds of individual infected cells to reveal the complexity of viral populations. We obtained these genotypes in parallel with host cell transcriptome information, enabling us to link host cell transcriptional phenotypes to the genetic structures underlying virus adaptation.
Project description:Purpose: Circular RNA sequencing was used to find out differentially expressed CircRNAs between P0 generation samples and P2 generation samples. Method: chondrocyte CircRNAs profiles of P0 generation samples and P2 generation samples were analyzed.
Project description:The Core Binding Factor (CBF) protein RUNX1 is a master regulator of definitive hematopoiesis, crucial for hematopoietic stem cell (HSC) emergence during ontogeny, which also plays vital roles in adult mice, in regulating the correct specification of numerous blood lineages. Akin to the other mammalian Runx genes, Runx1 has two promoters P1 (distal) and P2 (proximal) which generate distinct protein isoforms. The activities and specific relevance of these two promoters in adult hematopoiesis remain to be fully elucidated. Utilizing a dual reporter model we demonstrate here that the distal P1 promoter is broadly active in adult hematopoietic stem and progenitor cell (HSPC) populations. By contrast the activity of the proximal P2 promoter is more restricted and its upregulation, in both the immature Lineage- Sca1high cKithigh (LSK) and bipotential Pre-Megakaryocytic/Erythroid Progenitor (PreMegE) populations, coincides with a loss of erythroid specification. Accordingly the PreMegE population can be prospectively separated into âpro-erythroidâ and âpro-megakaryocyteâ populations based on Runx1 P2 activity. Comparative gene expression analyses between Runx1 P2+ and P2- populations indicated that the level of CD34 expression could substitute for P2 activity to distinguish these two cell populations in wild type (WT) bone marrow (BM). Prospective isolation of these two populations will provide the opportunity to further investigate and define the molecular mechanisms involved in megakaryocytic/erythroid (Mk/Ery) cell fate decisions. mRNA profiles of wild type (WT), Runx1 P2-hCD4+ (P2+) and Runx1 P2-hCD4- (P2-) Bone marrow Pre-Megakryocyte/Erythroid (PreMegE) progenitors were generated from young adult (12-16 weeks) mice by deep sequencing, in triplicate, using Illumina NextSeq 500.
Project description:We have identified two vessel forming mesenchymal stromal cell populations. Population 1 (P1) is CD45-Ter119-Tie2+PDGFRa-CD31+CD105highSca1low and gives rise to stunted vessels (incomplete tubular structures) in a transplant setting, and Population 2 (P2) which is CD45-Ter119-Tie2+PDGFRa+CD31-CD105lowSca1high and forms stunted vessels and fat. To test the homogeneity and signal interactions of these populations we performed 10X single cell RNA-sequencing on them.
Project description:Hepatocyte Nuclear Factor 4α (HNF4α), master regulator of hepatocyte differentiation, is regulated by two promoters (P1 and P2). P1-HNF4α but not P2-HNF4α is expressed in normal adult liver in fed conditions. Both P1- and P2-HNF4α are expressed in fetal liver. P2-HNF4α expression is increased in fasted conditions, high fat diet, alcoholic liver and liver cancer. To determine the target genes of the P1- and P2-HNF4α isoforms, we compared P2-HNF4α-expressing exon swap mice (a7HMZ) to wildtype (WT) male mice. Liver ChIP-seq samples were taken at 10:30 AM (ZT 3.5)
Project description:Viruses can disperse collectively using extracellular vesicles and other types of multi-virion structures. It has been hypothesized that, by increasing the cellular multiplicity of infection, this dispersal mode may favor cooperation among viral genomes. However, the spread of defective variants that function as social cheaters could also be promoted. To better understand the nature of virions present in vesicles, here we examine the genetic diversity harboured by vesicles of coxsackievirus B3 (CVB3), a model enterovirus. Our results confirm that CVB3 vesicles contain multiple infectious particles. However, we also find that virus variants coinfecting a cell typically allocate their progenies into different vesicles, precluding long-term interactions among them. Furthermore, Illumina sequencing indicates that dispersal through vesicles does not increase viral population genetic diversity appreciably. We conclude that vesicles enable the co-dispersal of groups of virions, but that these groups are highly related since they share the same parental genome. This should restrict the evolution of defective viruses, but also makes cooperation among different viral genetic variants unlikely. Our results are in line with a fundamental tenet of social evolution theory, according to which the evolution of cooperation typically requires that the interacting partners are genetically related.