RNA sequencing of E15.5 isolated progenitors cells from wildtype and Zmiz1-KO cortex
Purpose: Zinc Finger MIZ-Type Containing 1 (Zmiz1) is a member of the PIAS family of protein and function as a transcriptional coactivator of Notch, Androgen Receptor (AR), p53, Estrogen Receptor (ER), and Smad3/4 . Despite Zmiz1 mutations association with neurodevelopmental disorders such as ASD, ADHD, and intellectual disability, its role in physiological and pathological neurodevelopment  is significantly unknown. Here, we use murine model to knockout Zmiz1 using Emx1Cre and performed basal progenitors isolation at E15.5 and RNA sequencing to profile transcriptional changes upon Zmiz1 deletion. Timed pregnancy was carried out. Mice was euthanized using CO2 and embryos brain were extracted at E15.5. Cortex was dissected and dissociated into single cell suspension using Neural Tissue Dissociation Kit (P) (Miltenyi Biotec, 130-092-628) following manufacture instruction. Intermediate progenitors were magnetically selected and isolated using Anti-Prominin-1 MicroBeads, mouse (Miltenyi Biotec, 130-092-333). Total RNA was extracted from wildtype and Zmiz1-KO samples. RNA concentration and RNA integrity number were determined. RNA library was prepared, quantified, and verified using TruSeq RNA Library Prep Kit v2, Qubit dsDNA High Sensitivity Assay kit and Bioanalyzer DNA1000 assay kit respectively. Verified samples were sequenced using the NextSeq1000/2000 P2 Reagents (200 Cycles) v3 on a Nextseq1000/2000. RNA-seq data analysis was performed using illumina BaseSpace Sequence Hub. Briefly, sequenced reads were aligned to mouse (mm10) reference genome with RNA-Seq alignment tool (STAR aligner) and differentially expressed genes (DEG) were determined using the RNA-Seq Differential Expression tool (version 1.0.1). Results: We found 261 differentially expressed genes of which 67 genes were upregulated while 194 genes were downregulated. Downregulated genes were enriched in biological processes such as central nervous system differentiation, axon development, neuron migration, progenitors proliferation, etc. Conclusions: We assessed cortical intermediate progenitors transcriptional profile potentially regulated by Zmiz1. Overall design: E15.5 isolated WT and Zmiz1-KO progenitors RNA sequencing