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Quantitative trait locus mapping for interleukin-1β release after inflammasome priming and activation was performed on bone-marrow-derived macrophages (BMDM) from an AKRxDBA/2 mouse strain intercross. The strongest associated locus mapped very close to the  Pycard  gene on chromosome 7, which codes for the inflammasome adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC). The DBA/2 and AKR  Pycard  genes only differ at a single-nucleotide polymorphism (SNP) in their 3’ untranslated region (UTR). DBA/2 vs. AKR BMDM had increased levels of  Pycard  mRNA expression and ASC protein, and increased inflammasome speck formation, which was associated with increased  Pycard  mRNA stability without an increased transcription rate. CRISPR/Cas9 gene editing was performed on DBA/2 embryonic stem cells to change the  Pycard  3’UTR SNP from the DBA/2 to the AKR allele. This single base change significantly reduced  Pycard  expression and inflammasome activity after cells were differentiated into macrophages due to reduced  Pycard  mRNA stability.

Genetic variant in 3’ untranslated region of the mouse pycard gene regulates inflammasome activity