RNA sequencing data from the prefrontal cortex and hippocampus of male (12 weeks old) hemizyguous CAG-HERV-W-env mice and wild-type controls
Description
RNA sequencing data from the prefrontal cortex (PFC) and hippocampus (HIPP) of male (12 weeks old) hemizyguous CAGHERV-Wenv mice ( C57BL6/J;129P2/Ola-Hprt mice; n = 3) relative to wild-type ( n = 3) littermates. Total RNA was extracted from prefrontal and hippocampal samples using the SPLIT RNA extraction kit (Lexogen, Austria) following the manufacturer’s recommendations and was sent to the Functional Genomics Center in Zurich (FGCZ) for quality control and RNA sequencing. The quality of the isolated RNA was determined with a Fragment Analyzer (Agilent, Santa Clara, California, USA). Only those samples with a 260 nm/280 nm ratio between 1.8–2.1, a 28S/18S ratio within 1.5–2, and RIN (>8) values qualified for a Poly-A enrichment strategy in order to generate the sequencing libraries applying the TruSeq mRNA Stranded Library Prep Kit (Illumina, Inc, California, USA). After Poly-A selection using Oligo-dT beads the mRNA was reverse-transcribed into cDNA. The cDNA was fragmented, end-repaired and poly-adenylated before ligation of TruSeq UD Indices (IDT, Coralville, Iowa, USA). The quality and quantity of the amplified sequencing libraries were validated using a Fragment Analyzer SS NGS Fragment Kit (1–6000 bp) (Agilent, Waldbronn, Germany). The equimolar pool of the samples was spiked into a NovaSeq6000 run targeting ~15M reads per sample on a S1 FlowCell (Novaseq S1 Reagent Kit, 100 cycles, Illumina, Inc, California, USA). Reads were quality-checked with FastQC. Sequencing adapters were removed with Trimmomatic and aligned to the reference genome and transcriptome of Mus Musculus (GENCODE, GRCm38,p5) with STAR v2.7.3. Distribution of the reads across genomic isoform expression was quantified using the R package GenomicRanges from Bioconductor Version 3.10. Minimum mapping quality, as well as minimum feature overlaps, was set to 10. Multi-overlaps were allowed. Differentially expressed genes (DEGs) were identified using the R package edgeR from Bioconductor Version 3.10, using a generalized linear model (glm) regression, a quasi-likelihood (QL) differential expression test and the trimmed means of M-values (TMM) normalization.
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Funding
- Swiss National Science Foundation
- Experimental Investigations of Human Endogenous Retrovirus Type W (HERV-W) in Relation to Psychiatric and Neurological Disorders 320030-231937
Dates
- Created
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2024-11-18