Functional Genomic Analysis of a RUNX3 Polymorphism Associated With Ankylosing Spondylitis.

ABSTRACT:

Objective

To investigate the functional consequences of the single-nucleotide polymorphism rs4648889 in a putative enhancer upstream of the RUNX3 promoter associated with susceptibility to ankylosing spondylitis (AS).

Methods

Using nuclear extracts from Jurkat cells and primary human CD8+ T cells, the effects of rs4648889 on allele-specific transcription factor (TF) binding were investigated by DNA pull-down assay and quantitative mass spectrometry (qMS), with validation by electrophoretic mobility shift assay (EMSA), Western blotting of the pulled-down eluates, and chromatin immunoprecipitation (ChIP)-quantitative polymerase chain reaction (qPCR) analysis. Further functional effects were tested by small interfering RNA knockdown of the gene for interferon regulatory factor

SUBMITTER: Vecellio M 

PROVIDER: S-EPMC8251554 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature