Project description:Mar1 deletion and RNA enrichment in Cryptococcus neoformans: pilot data for a high-throughput sequencing course. The goal of this project was to generate pilot data in preparation for a summer course on high-throughput sequencing where participants prepared their own RNA-Seq libraries and analyzed the resulting data. This pilot experiment addressed two questions: 1. Does this experimental system (Cryptococcus neoformans H99 wildtype and mar1 deletion mutant grown in YPD and tissue culture media) provide a good dataset for course participants to analyze. 2. Which rRNA depletion method is best to use in the wetlab component of the course. This data was generated in preparation for the intensive summer course on high-throughput sequencing, funded by NIH grant 5R25EB023928-03 "A hands-on, integrative next-generation sequencing course: design, experiment, and analysis".
Project description:The biological function and disease association of human endogenous retroviral (HERV) elements remains largely elusive. We addressed the physiological role of HERV-K(HML-2) in neuronal differentiation by manipulating HERV-K(HML-2) expression levels. We used CRISPR engineering to activate or repress HERV-K(HML-2) and demonstrate that elevated HERV-K(HML-2) transcription is detrimental for development, functionality and growth of cortical neurons. Effects are cell-type specific, as dopaminergic neurons were unaffected. We further show that layer formation is altered during forebrain organoid formation following activation of HERV-K(HML-2) transcription. HERV-K(HML-2) transcriptional activation concurrently elevated Neurotrophic Tyrosine Receptor Kinase 3 (NTRK3) expression along with other neurodegeneration-related genes. Direct transcriptional activation of NTRK3 resembled the HERV-K(HML-2) activation phenotype. Intriguingly, reduction of NTRK3 levels in HERV-K(HML-2)-activated cortical neurons restored differentiated cortical neurons. Hence, our findings unravel a unique cell type-specific mechanism of HERV-K(HML-2) during cortical neuronal differentiation.
Project description:Elevated transcript expression of the endogenous retrovirus family HERV-K (HML-2) is seen in the majority of breast cancers, although the identity of the individual loci contributing to this expression as well as their mechanism of activation is unclear. Using high-throughput next-generation sequencing techniques optimized for the capture of HML-2 expression, we produced a complete profile of the HML-2 transcriptome before and after human mammary epithelial cell transformation.
Project description:A pilot experiment for an RNA-seq dataset (SRP003234). This experiment compares AP1 domain-specific translating RNA with total flower translating mRNA.
Project description:We report nucleosome positions by using native Mnase-seq. The results strongly suggest that nucleosome positions don't change at the heterochromatic locus HMR upon removal of adjacent nucleosomes via deletion of nucleosomal DNA. Positions of nucleosomes at HML are also quantified.
2019-09-06 | GSE136897 | GEO
Project description:Target enrichment sequencing of HERV-K (HML-2)
Project description:Drosophila carrying Hml-SwitchGal4 were crossed to different uas-constructs (uas-eGFP, uas-PGRP-LC, uas-hop, uas-tkvQD). 3rd instar larvae were either kept as control or SwitchGal4 was activated by hormone feeding for 24h. Afterwards plasmatocytes were extracted and subjected to RNA-seq.
Project description:We identified target genes for NHR-25 by ChIP-seq at L1 stage of C. elegans. Transcription factor genes were tagged with GFP and their expression examined at L1 stage. Since there are no direct target genes known for NHR-25 that can be used for assessment of enrichment efficiency by quantitative PCR (qPCR), we chose to repeat ChIP-seq experiment of another GFP tagged transcription factor, PHA-4 for which the ChIP-seq was performed during a pilot experiment of modENCODE project using the same transgenic strain and antibody (a gift from Tony Hyman lab).
