RNA-seq analysis of parental (wild type) and dcl3 mutant lines of Chlamydomonas reinhardtii
ABSTRACT: DCL3 is the main processor of miRNA precursors in Chlamydomonas reinhardtii. Because of that, and in order identify mRNA targets of miRNA-guided slicer complexes, we used RNAseq of the transcriptome in dcl3 mutant and parental lines.
We describe here a forward genetic screen to investigate the biogenesis, mode of action, and biological function of miRNA-mediated RNA silencing in the model algal species,Chlamydomonas reinhardtii Among the mutants from this screen, there were three at Dicer-like 3 that failed to produce both miRNAs and siRNAs and others affecting diverse post-biogenesis stages of miRNA-mediated silencing. The DCL3-dependent siRNAs fell into several classes including transposon- and repeat-derived siRNAs as in ...[more]
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: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:Here, we report on the transcriptome of the organelles of the micro-alga, Chlamydomonas reinhardtii, sampled under a number of different conditions. The preparation of the RNA-Seq libraries and their analysis were performed using protocols optimized for organellar transcripts. Samples include growth in media +/– Fe, growth in media +/– Cu, diurnal growth samples collected in dark and light, and the sexual cycle. Overall design: RNA-Seq libraries were prepared from Chlamydomonas reinhardtii sampled under 12 different conditions including: diurnal growth in light, diurnal growth in dark, +Fe, –Fe, +Cu, –Cu, mt+ vegetative, mt– vegetative, mt+ gamete, mt– gamete, zygote, and post-germination.
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
Project description:We analysed global gene expression changes in Chlamydomonas reinhardtii in response to 1h UV-B, applied at the same low level that was seen to promote subsequent UV-B stress tolerance, in order to elucidate the transcriptional reprogramming that leads to UV-B acclimation. mRNA profiles generated by deep sequencing from triplicate replicate Chlamydomonas reinhardtii samples sourced from independent cultures either protected from UV-B or exposed to 1h acclimation-level UV-B.
Project description:This SuperSeries is composed of the following subset Series: GSE20859: Change in gene expression in Chlamydomonas reinhardtii upon heat shock GSE20860: Change in gene expression in Chlamydomonas reinhardtii upon feeding with hemin and Mg-protoporphyrin Refer to individual Series
Project description:endogenous small RNAs from Chlamydomonas reinhardtii strain J3(mt-) vegetative cells Keywords: High throughput 454 small RNA sequencing Overall design: Size fractionated small RNA from total RNA extracts was ligated to adapters, purified again and reverse transcribed. After PCR amplification the sample was subjected to 454 high throughput pyrosequencing. Please see www.454.com for details of the sequencing technology.
Project description:This data was generated to identify the molecular pathways responsible for nitrous oxide synthesis by the green algae Chlamydomonas reinhardtii, when supplied with nitrite under aerobic conditions (oxia). RNA samples were collected at three time points, 15 min, 3 hours, and 24 hours after the start of the experiment. The control and treatment groups were grown under the same conditions, except treatment group was supplied with 10mM nitrite at time 0. Illumina TruSeq stranded RNA libraries were synthesised from the resulting RNA before sequencing on a HiSeq2500 (125bp). The resulting sequence run generated 241,151,809 paired-end 125bp reads, of which 200,946,839 remained following quality filtering. The short data was mapped to the published genome and read counts were generated with HT-Seq count with the default settings. The raw read count data was analysed by DESeq2 in order to identify genes differentially expressed during nitrous oxide production. Overall design: Stranded RNA-seq libaries were synthesised from RNA collect at 15 minutes, 3 hours, and 24 hours after addition of 10 mM nitrite to the treatment group of Chlamydomonas reinhardtii. Controls were grown under identical conditions but not supplied with nitrite. Three growth flask were used for the control and treatment group to provide triplicates.
Project description:Zn-limited cells accumulate Cu in intracellular structures in a reversable manner. We used RNAseq analysis to monitor gene expression in Zn-limited cells, and during a time course (0 - 24 h) after Zn resupply. Overall design: Comparison of Chlamydomonas reinhardtii gene expression when Zn deficient cells are resupplied with 2.5 µM Zn. Time points 0 (before resupply), 1.5, 3, 4.5, 12, and 24 hours after resupply. Four biological replicates per condition.