Transcription factor network analysis identifies REST/NRSF as an intrinsic regulator of CNS regeneration

The goal of this study is to use RNA-seq to investigate the molecular mechanisms by which REST inhibition regulates corticalspinal axon regeneration. To achieve this goal, cortical motor neurons from adult wild-type (WT) and REST conditional knockout (cKO) were purified at 1, 3, 7 days following spinal cord injury (SCI) or sham operation. RNA-seq libraries for cortical motor neurons were prepared with the QuantSeq mRNA-Seq library prep kit FWD for Illumina (Lexogen) and sequenced using HiSeq4000 (Illumina). Sequenced reads were mapped to reference genome (mm10) using STAR, and count level data were quantifed using Salmon. Differential gene expression analysis was performed using limma voom. mRNA profiles were performed in cortical motor neurons from wild-type or REST cKO mice that were recovered 1, 3, 7 days following SCI or sham operation. Each group has 3-4 replicates with ~1000 cells in each replicate.