Bulk RNA sequencing analysis of a patient with an immunodeficiency caused by a novel homozygous missense variant in HYOU1
Bulk RNA sequencing analysis of a patient with an immunodeficiency caused by a novel homozygous missense variant in HYOU1
For PBMC: Standard venopuncture followed by ficoll extraction of PBMC For BM:  For HDF:Human dermal fibroblast (HDF) isolation of the proband was performed within three days of the skin biopsies. The biopsy was transferred to a sterile petri dish. Dermis and hypodermis were removed and discarded. Epidermis was cut into small pieces and placed in 25 cm2 flasks. Culture media (Dulbecco's Modified Eagle Medium (10566016, Gibco™, Thermo Fisher Scientific, Waltham, MA, USA) (DMEM) supplemented with 10% inactivated foetal bovine serum (FBS), 40 U/ml penicillin, and 50 mg/ml streptomycin was added to cover the surface after 15 minutes. The media was changed after 24 hours. The cells were passed at 80% confluency using trypsinization. Control HDFs were purchased from Promocell (Heidelberg, Germany). RNA was extracted using the RNeasy Mini Kit (74106, Qiagen, Hilden, Germany).  SMARTer® Stranded Total RNA-Seq Kit v2 (Takara, Otsu, Shiga, Japan)