Human TBK1 Deficiency Leads to Autoinflammation Driven by TNF-Induced Cell Death - Bulk RNAseq

Published: 23 June 2021| Version 1 | DOI: 10.17632/648zntv5dx.1
Contributor:
Justin Taft

Description

Bulk RNA sequencing of hTERT-immortalized dermal fibroblasts derived from two TBK1-deficient patients and three HCs following a 24 hr stimulation with 10 ug/mL poly(I:C) with and without Lipofectamine 2000.

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Steps to reproduce

hTERT-immortalized dermal fibroblasts were plated at 75,000 cells/well in a 24-well plate day before in 500 uL DMEM (10% FBS, 1% GlutaMAX, 1% P/S). Cells were treated as indicated with either DMEM alone, L2K, 10 ug/mL pIC, or 10 ug/mL pIC+L2K for 24 hrs. After stimulation, cells were lysed and RNA was extracted with the RNeasy RNA Isolation Kit (Qiagen) and treated with RNase-Free DNase (Qiagen) to remove DNA. The libraries were prepared with the TruSeq RNA Sample Prep Kit v2 (Illumina). FASTQ files from the NextSeq 550 (Illumina) were uploaded to BaseSpace and analyzed using the RNA-Seq Alignment (STAR) and RNA-Seq Differential Expression (DESeq2) modules.

Institutions

Icahn School of Medicine at Mount Sinai

Categories

Viral Disease, Autoinflammatory Disease

License