Project description:endogenous small RNAs from Chlamydomonas reinhardtii strain J3(mt-) vegetative cells Keywords: High throughput 454 small RNA sequencing
Project description:RNA populations in Chlamydomonas reinhardtii Keywords: Highly parallel pyrosequencing Small RNAs were prepared from Chlamydomonas reinhardtii total extracts,ligated to a 3' adaptor and a 5' acceptor sequentially, and then RT-PCR amplified. PCR products were reamplified using a pair of 454 cloning primers and provided to 454 Life Sciences (Branford, CT) for sequencing. For technical details, see Tao Zhao, Guanglin Li, Shijun Mi, Shan Li, Gregory J. Hannon, Xiu-Jie Wang, and Yijun Qi. 2007. A Complex System of Small RNAs in the Unicellular Green Alga Chlamydomonas reinhardtii. Genes & Development
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from cultures of Chlamydomonas reinhardtii (in control, phosphate starvation and sulphate starvation conditions). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the genome under study. Small RNA libraries were derived from cultures of Chlamydomonas reinhardtii in control, phosphate starvation and sulphate starvation conditions. Total RNA was isolated using the TriReagent (Molecular Research Center), and submitted to Illumina (Hayward, CA, http://www.illumina.com) for small RNA library construction using approaches described in (Lu et al., 2007) with minor modifications. The small RNA libraries were sequenced with the Sequencing-By-Synthesis (SBS) technology by Illumina. PERL scripts were designed to remove the adapter sequences and determine the abundance of each distinct small RNA. We thank Kan Nobuta and Gayathri Mahalingam for assistance with the computational methods.
Project description:The metabolites derived from microalgae have been attributed with various nutritional and medicinal properties. Therefore, our study aimed to investigate the potential beneficial effects of Chlamydomonas reinhardtii (red), a type of microalgae, in individuals with type 2 diabetes mellitus (T2DM). Mice were fed on high-fat diet and injected with a low dose of streptozotocin to induce T2DM. The diabetic mice were orally treated with either 1% sodium carboxymethylcellulose or Chlamydomonas reinhardtii (red) at doses of 1, 2, or 3 g/kg BW/day for a duration of 4 weeks. The liver sections were subjected to hematoxylin and eosin staining as well as oil red staining for the detection of pathological changes and lipid deposition, respectively. Inflammatory factors in serum were quantified using ELISA kits, while commercial kits were employed to assess oxidative stress-related indicators. Gene expression in liver was analysed by RNA-seq. The results revealed that Chlamydomonas reinhardtii (red) significantly ameliorated fasting blood glucose levels, body weight, triglyceride, and low density lipoprotein cholesterin, while also enhancing oral glucose tolerance and insulin sensitivity. In pathological analysis, Chlamydomonas reinhardtii (red) significantly improved lipid deposition and hepatic tissue damage. Furthermore, Chlamydomonas reinhardtii (red) could obviously decreased the protein expression of G-6-Pase and PEPCK, and regulated the SOCS2/JAK2/STAT5 signaling pathway. Transcriptomic analysis indicated that a total of 972 significantly differentially expressed genes in diabetic mice treated with Chlamydomonas reinhardtii (red). KEGG analyses revealed that lipid and atherosclerosis, MAPK signaling pathway, B cell receptor signaling pathway, TNF signaling pathway, NOD-like receptor signaling pathway, Toll-like receptor signaling pathway, PI3K-Akt signaling pathway were involved in Chlamydomonas reinhardtii (red) modulated process. Therefore, the continuous consumption of Chlamydomonas reinhardtii (red) may have anti-T2DM effects through the inhibition of gluconeogenesis, thus offering a promising alternative for T2DM patient.
Project description:DCL3 appeared several times in a forward genetic screen meant to isolate genes involved in miRNA-mediated RNA silencing in Chlamydomonas reinhardtii. Because of that, and in order to define the role of DCL3 in the processing of small RNAs on a genome-wide basis, we sequence the small RNA transcriptome of dcl3 mutant and parental lines.
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from cultures of Chlamydomonas reinhardtii (in control, phosphate starvation and sulphate starvation conditions). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the genome under study.
Project description:Chlamydomonas reinhardtii exposed to various concentrations of silver For this experiment,C. reinhardtii were exposed to (4) different concentrations of silver, as biological triplicates