Coilin Mutation in Zebrafish Impairs the Innate Immune Zebrafish
Coilin is a nuclear protein known for its roles in assembling the Cajal Body and participating in the biogenesis of ribonucleoproteins. We have uncovered novel functions for coilin in promoting microRNA biogenesis and the phosphorylation/SUMOylation of various proteins, through which we have observed a role for coilin in responding to stress signals. Coilin has also been found to participate in the stress response in plants through the regulated expression of immunity genes and activation of defense mechanisms. In this study, we further investigated this role of coilin in regulating the immune response by applying transcriptomic analyses to examine the response to lipopolysaccharide-mediated stress in coilin-depleted human cells and coilin mutant zebrafish embryos. Our results indicate a vital function for coilin in regulating the expression of immunity-related genes and establish a conserved role for vertebrate coilin in the promotion of the innate immune response across humans and zebrafish. Overall design: RNA was isolated by TRIzol Reagent (Invitrogen Life Technologies, Grand Island, NY, USA) and purified with RNeasy Mini Kit (QIAGEN; Hilden, Germany) according to manufacturer's instructions. RNAseq was used to analyze differential expression between the following groups. RNA was evaluated for quality (Qubit fluorimeter) and fidelity using Qiagen QIAxcel Advanced System, with all sample demonstrating and RIS > 8. Libraries were generated with TruSeq stranded mRNA kit (Illumina; San Diego, CA, USA), indexed with TruSeq RNA CD indexes (Illumina; San Diego, CA, USA), and sequenced with NextSeq 2000 P3 (PE200) flow cell (Illumina; San Diego, CA, USA)