Project description:Male skeletal muscles are generally faster and have a higher maximum power output than female muscles. Conversely, during repeated contractions female muscles sre generally more fatigue resistant and recover faster. we hypothesized that estrogen receptor beta is involved in this gender difference. The Affymetrix micorarray were used to deteced the differently expressed genes between male BERKO and WT mice in soleus muscles. Soleus muscles from 3 individual mouse of each genotype were used to perform the experiment.
Project description:Detecting strain-specific barcodes with mass spectrometry can facilitate the screening of genetically engineered bacterial libraries. Here, we introduce intact protein barcoding, a method to measure protein-based library barcodes and metabolites using flow-injection mass spectrometry (FI-MS). Protein barcodes are based on ubiquitin with N-terminal tags of six amino acids. We demonstrate that FI-MS detects intact ubiquitin proteins and identifies the mass of N-terminal barcodes. In the same analysis, we measured relative concentrations of primary metabolites. We constructed 6 ubiquitin-barcoded CRISPRi strains targeting metabolic enzymes, and analyzed their metabolic profiles and ubiquitin barcodes. FI-MS detected barcodes and distinct metabolome changes in CRISPRi-targeted pathways. We demonstrate the scalability of intact protein barcoding by measuring 132 ubiquitin barcodes in microtiter plates. These results show that intact protein barcoding enables fast and simultaneous detection of library barcodes and intracellular metabolites, opening up new possibilities for mass spectrometry-based barcoding.
Project description:Assessment of zooplankton diversity integrating microscopy and multigene high-throughput sequencing in the Bay of Malaga (SW Mediterranean). Metagenome
Project description:Male skeletal muscles are generally faster and have a higher maximum power output than female muscles. Conversely, during repeated contractions female muscles sre generally more fatigue resistant and recover faster. we hypothesized that estrogen receptor beta is involved in this gender difference. The Affymetrix micorarray were used to deteced the differently expressed genes between male BERKO and WT mice in soleus muscles. Soleus muscles from 3 individual mouse of each genotype were used to perform the experiment. Keywords: repeat sample
Project description:LNPs have been demonstrated to hold great promise for the clinical advancement of RNA therapeutics. Continued exploration of LNPs for application in new disease areas requires identification and optimisation of leads in a high throughput way. Currently available high throughput in vivo screening platforms are well suited to screen for cellular uptake but less so for functional cargo delivery. We report on a platform which measures functional delivery of LNPs using unique peptide ‘barcodes’. We describe the design and selection of the peptide barcodes and the evaluation of these for the screening of LNPs. We show that proteomic analysis of peptide barcodes correlates with quantification and efficacy of barcoded reporter proteins both in vitro and in vivo and, that the ranking of selected LNPs using peptide barcodes in a pool correlates with ranking using alternative methods in groups of animals treated with individual LNPs. We show that this system is sensitive, selective, and capable of reducing the size of an in vivo study by screening up to 10 unique formulations in a single pool, thus accelerating the discovery of new technologies for mRNA delivery.
Project description:Pluripotency of mammalian stem cells is stabilized at different status depending upon the extracellular stimuli of their environment. Now, there is a consensus that mouse embryonic stem cells (mESCs) and iPSCs (miPSCs) represent undifferentiated status very close to ground-state of mammalian development when cultured in optimal condition. Since they have not been activated to commit into any lineages, they are stated as “naïve-state stem cells”. In contrast, human ESCs (hESCs), hiPSCs, and mEpiSCs are considered to possess partially differentiated characteristics compared with mESCs and miPSCs. Thus, they are stated as “primed-state stem cells”. Investigation of the prerequisites for establishment of the naïve-state is significantly important to fully understand mammalian development and to extrapolate the technologies developed with mouse ES/iPS cells for human. We used expression data to understand the gene expression profile and identified distinct characteristics on pluripotent state.
Project description:Pluripotency of mammalian stem cells is stabilized at different status depending upon the extracellular stimuli of their environment. Now, there is a consensus that mouse embryonic stem cells (mESCs) and iPSCs (miPSCs) represent undifferentiated status very close to ground-state of mammalian development when cultured in optimal condition. Since they have not been activated to commit into any lineages, they are stated as “naïve-state stem cells”. In contrast, human ESCs (hESCs), hiPSCs, and mEpiSCs are considered to possess partially differentiated characteristics compared with mESCs and miPSCs. Thus, they are stated as “primed-state stem cells”. Investigation of the prerequisites for establishment of the naïve-state is significantly important to fully understand mammalian development and to extrapolate the technologies developed with mouse ES/iPS cells for human. We used expression data to understand the gene expression profile and identified distinct characteristics on pluripotent state.