Cynoglossus semilaevis testis cells RNA-Seq after hnrnpk/tox2 overexpression
This study performed transcriptome sequencing on testicular cells of the Chinese tongue sole (Cynoglossus semilaevis) to investigate the regulatory roles of the hnrnpk and tox2 genes. Following overexpression of hnrnpk and tox2, and using an empty vector as a negative control, total RNA was extracted and strand-specific cDNA libraries were constructed. Sequencing was conducted on the Illumina NovaSeq 6000 platform with a paired-end 150 bp strategy. The raw reads were filtered to remove adapters and low-quality sequences. The clean reads were then aligned to the C. semilaevis reference genome, and transcript assembly and quantification were performed. Differential expression analysis between the overexpression groups and the control group was conducted to identify genes with significant expression changes (|fold change| >= 2, FDR < 0.05). Subsequent analyses included Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment for the differentially expressed genes, as well as gene set enrichment analysis (GSEA) and alternative splicing event detection.