For the study of mRNA decay rates, transcription was inhibited with ActinomycinD, and RNA splicing with Sinefungin, at different time points, in the Matthews lab. rRNA depleted RNA was extracted from each of the samples in the Clayton lab, and sent for deep sequencing at the BioQuant facility in Heidelberg
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Projects: SilicoTryp
Institutions: University of Edinburgh
I am a Postdoc at Keith Matthews lab in the Institute of Immunology and Infection Research, Edinburgh University. As part of the SilicoTryp project we are in charge of performing Targeted disruption and Overexpression of critical enzymes of Trypanosoma brucei redox metabolism enzymes and developmental perturbations to provide part of the necessary data for the construction of the model. Also generate consistent samples, so that data can be integrated and quantification results are guarateed to ...
SysMO is a European transnational funding and research initiative on "Systems Biology of Microorganisms".
The goal pursued by SysMO was to record and describe the dynamic molecular processes going on in unicellular microorganisms in a comprehensive way and to present these processes in the form of computerized mathematical models.
Systems biology will raise biomedical and biotechnological research to a new quality level and contribute markedly to progress in understanding. Pooling European research ...
Projects: BaCell-SysMO, COSMIC, SUMO, KOSMOBAC, SysMO-LAB, PSYSMO, SCaRAB, MOSES, TRANSLUCENT, STREAM, SulfoSys, SysMO DB, SysMO Funders, SilicoTryp, Noisy-Strep
Web page: http://sysmo.net/
The SilicoTryp project aims at the creation of a “Silicon Trypanosome”, a comprehensive, experiment-based, multi-scale mathematical model of trypanosome physiology. Trypanosomes are blood-stream parasites transmitted by tsetse flies; they cause African sleeping sickness in humans and livestock. Currently available drugs have severe side effects, and the parasites are rapidly developing resistance. In this project, we collect a wide range of new experimental data on the parasite in its various ...
Programme: SysMO
Public web page: http://silicotryp.ibls.gla.ac.uk/wiki/Main_Page
Organisms: Trypanosoma brucei
Aim: To provide quantitative data that will allow modeling of gene expression for all enzymes of redox metabolism and the pentose phosphate pathway. Modeling will be used to predict enzyme levels based on the integration of an RNA degradation model with translation and protein degradation rates.
Plan: The amounts of a protein in a cell can be determined by the rates of transcription, mRNA processing, translation, mRNA turnover and protein degradation. In trypanosomes analysis is simpler because ...
Submitter: Abeer Fadda
Studies: Determination and integration of abundance, processing efficiency, and d...
Assays: Modelling the gene expression cascade with length-dependent processes, mRNA decay assay, pre-mRNA processing rate
Snapshots: No snapshots
Since over 40 enzymes will be investigated for their mRNA abundance, processing, and degradation kinetics, the less tedious and more accurate Next Generation Sequencing of the entire mRNA repertoire of the cell is employed. To optimise the proportion of useful sequence, while including RNA fragments that are products of of degradation, rRNA is depleted using the eukaryotic Ribominus kit (Ambion). Two biological replicates are treated with Sinefungin and Actinomycin D to inhibit RNA processing and ...
Submitter: Abeer Fadda
Investigation: Gene expression in Trypanosoma brucei
Assays: Modelling the gene expression cascade with length-dependent processes, mRNA decay assay, pre-mRNA processing rate
Snapshots: No snapshots
This assay is designed to measure the decay kinetics of mRNA in T. brucei blood forms. T. brucei lacks the canonical transcriptional regulation employed by other eukaryotes through transcription factors, and relies almost entirely on regulation of mRNA decay and further downstream steps in order to control gene expression. 3 replicates of 8 time points were taken to measure mRNA abundance in the cell using RNA-seq. The first time point was fot WT, untreated cells; the second was 5 min after the ...
Submitter: Abeer Fadda
Assay type: Transcriptomics
Technology type: Technology Type
Investigation: Gene expression in Trypanosoma brucei
Organisms: Trypanosoma brucei : 2T1 (wild-type / wild-type)
SOPs: Sample preparation for mRNA decay study
Data files: Half life values for enzymes of redox and polya..., mRNA decay, mRNA half-lives
Snapshots: No snapshots