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FACS Sequencing Analysis of DCX+ cells in the DG of Wild Type and Fmr1-/y; Dcx-DsRed mice

ABSTRACT: Purpose: The goals of this study are to investigate the molecular mechanism underlying FMRP-regulated maturation of newborn neurons Methods: DG tissue were isolated from 6-7 weeks old Fmr1-/y; Dcx-DsRed mice and their WT littermates. All cell populations were isolated into single cells using a Becton Dickinson FACS Aria II contained in a Biosafety Carbinet using 20 psi pressure and 100-μm nozzle aperture. 10,000 total alive or Dcx-DsRed+ alive Cells were collected directly in Trizol. Total RNA from the sorted cell was isolated using the Direct-zol™ RNA MiniPrep Kit. Strand-specific, poly(A) selected cDNA libraries were generated using Nugen Ovation® Ultralow Library Systems (Illumina) according to the manufacturer’s protocol. Cluster generation and high-throughput sequencing were performed on a HiSeq 2500 (Illumina), using the paired-end 100 bp protocol. Reads were aligned to the mouse genome GRCm38 with annotation from Gencode. Results: RNA isolated from DsRed+ cells were subjected to next generation sequencing and 519 differentially expressed (DE) genes were identified that showed significant changes (FDR-adjusted P < 0.05) between Fmr1 KO and WT cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE117111 | GEO | 2019/01/31

REPOSITORIES: GEO

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FACS Sequencing Analysis of DCX+ cells in the DG of Wild Type and Fmr1-/y; Dcx-DsRed mice

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