Loss of circRNAs from the crh-1 gene extends the mean lifespan in Caenorhabditis elegans.

Abstract

Accumulation of circular RNAs (circRNAs) during aging occurs on a genome‐wide level for multiple organisms, but its significance is unknown. Generating circRNA loss‐of‐function mutants is difficult because the vast majority of these RNAs are comprised of exons shared with protein‐coding mRNAs. In Caenorhabditis elegans, most circRNAs were previously found to accumulate during aging. Two of the most abundant, age‐accumulating circRNAs are generated from exon 4 of the crh‐1 gene (circ‐crh‐1). Here, we found that the biogenesis of circ‐crh‐1 was regulated by the double‐stranded RNA‐binding protein ADR‐1. We identified Reverse Complementary Match (RCM) sequences in introns flanking circ‐crh‐1. Using CRISPR‐Cas9, we deleted the downstream RCM and found that this completely eliminated expression of the circRNA without affecting linear mRNA expression from the crh‐1 gene. Remarkably, worms lacking circ‐crh‐1 exhibited a significantly longer mean lifespan. Lifespan was partially restored to wild type by expression of circ‐crh‐1 in neural tissues. Widespread transcriptome alterations in circ‐crh‐1 mutants were identified using RNA‐Seq. Moving forward, intronic RCM deletion using CRISPR should be a widely applicable method to identify lifespan‐regulating circRNAs in C. elegans.