Transcriptomic Analysis of Tityus confluens Venom Glands Uncovers a Diverse Array of Bioactive Components

Introduction: Scorpion envenomation is a growing public health concern in Brazil, driven by urbanization, poor sanitation, and environmental change. The medically important Tityus confluens resembles T. serrulatus and has been linked to severe and fatal envenomations in neighboring countries. Its venom contains bioactive peptides and enzymes that cause toxicity and physiological effects, while potentially offering pharmacological compounds targeting ion channels and other pathways. Transcriptomic and proteomic analyses can characterize venom, clarify toxicological mechanisms, and identify therapeutic candidates. Objectives: This study aimed to evaluate the transcriptomic profile of the venom glands of T. confluens by performing RNA sequencing and analyzing selected molecular structures in silico. Methodology: We collected scorpions by UV-assisted active search and dissected venom glands from ten specimens for RNA extraction and quality assessment. RNA was converted to cDNA, and Nextera XT libraries were sequenced on the NovaSeq X Plus (paired-end, 2 × 100 bp). After FastQC, reads were assembled de novo with Trinity. We annotated and classified transcripts using BLASTx, UniProt/Swiss-Prot, DIAMOND, Gene Ontology, InterProScan/PFAM, and KEGG. Selected transcripts were analyzed for expression, signal peptides, physicochemical properties, sequence similarity, and predicted structures using RSEM, bioinformatics tools, and AlphaFold. Results: Transcriptome analysis identified 65,537 assembled transcripts, of which 50,231 (76.74%) received functional annotations. Among these, 666 (1.33%) were associated with venom components, including ion channel-modulating toxins, antimicrobial and antihypertensive peptides, metalloproteinases, and phospholipases. Eleven of the most highly expressed transcripts encoded venom components, including the sodium channel toxin TcNaTx-1, the anionic antimicrobial peptide TcAP-1, and the metalloproteinase TcMMp-2. These components were further characterized using in silico structural and physicochemical analyses. Metalloproteinases accounted for 36% of venom-related transcripts, while antimicrobial peptides made up 10%, underscoring the venom's molecular diversity. Conclusions: These findings highlight promising molecules for further research into analgesic, antimicrobial, antitumor, and other pharmacological applications.