Transcriptional profile of homozygous inactivation of Nf1 in mature oligodendrocytes. Mus musculus

White matter abnormalities have been observed in Neurofiromatosis type 1 (NF1) patients and in mouse models of the disease, and disturbed myelin has been proposed to cause of neurological issues in NF1 patients. Analysis of the transcriptional changes following inactivation of Nf1 in mature oligodendrocytes, the cell type producing myelin, can give information on the molecular pathways affected in NF1. Homozygous inactivation of Nf1 was induced by Tamoxifen treatment of adult PlpCreER;Nf1flox/flox mice, 1 month later, RNA from the optic nerve was isolated and sequenced Overall design: We isolated optic nerves from adult control and Nf1 mutant animals 1 month after tamoxifen treatment and performed whole-genome RNA sequencing. Total RNA was isolated from optic nerves using Trizol reagent (Invitrogen) followed by chloroform extraction and purification by RNeasy kit (Qiagen). Integrity of RNA was evaluated by Agilent Bioanalyzer 2100 (RIN>8.5) and cDNA was amplified by Ovation system v2 (NuGEN). RNA sequencing (35-40 million sequencing reads) was performed using paired-end BP flow cell. RNA-Seq data was mapped to the reference genome mm10, and fold change and variance were calculated using DESeq2 package in R.