Project description:This project mainly aims to characterize the complex toxic components present in the venom of Trimeresurus malabaricus (Malabar pit viper). Since Trimeresurus malabaricus (Malabar pit viper) species are mainly inhabited to plantation crop areas, its envenomation is a serious threat to the human population thriving in these zones, especially to the plantation workers. Therefore, exploring the venom proteome of Malabar pit viper is decisive to develop and design new antivenom and therapeutics against its envenomation. As described in this study, applying various orthogonal separation strategies helped in dissecting venom constituents of Trimeresurus malabaricus and is the first comprehensive attempt in revealing the complex venom profile of Malabar pit viper through proteomics approaches incorporating multiple database searches. In order to achieve this the crude venom components were resolved on a 12% SDS page. Further each of the bands were subjected to in-gel trypsin digestion. The crude venom was also subjected to ion-exchange chromatography separation. The obtained fractions were subjected to in-solution trypsin digestion. All the digested peptides were then subjected to Q-TOF LC-MS/MS analysis.
Project description:Both single cell and bulk RNA sequencing was performed on expanding or differentiating snake venom gland organoids (from Aspidelaps Lubricus Cowlesi and Naja Nivea), or tissue (Aspidelaps Lubricus Cowlesi). Bulk RNA sequencing from the snake venom gland, liver and pancreas was performed to construct a de novo transcriptome using Trinity.
Project description:Diachasmimorpha longicaudata parasitoid wasps carry a symbiotic poxvirus, known as DlEPV, within the female wasp venom gland. We sequenced RNA from venom gland tissue to identify DlEPV orthologs for 3 conserved poxvirus core genes. The DlEPV ORFs identified from this transcriptome were used to design primers for downstream RT-qPCR analysis and RNAi knockdown experiments.
Project description:Background The generalist dipteran pupal parasitoid Nasonia vitripennis injects 79 venom peptides into the host before egg laying. This venom induces several important changes in the host, including developmental arrest, immunosuppression, and alterations to normal metabolism. It is hoped that diverse and potent bioactivities of N. vitripennis venom provide an opportunity for the design of novel acting drugs. However, currently very little is known about the individual functions of N. vitripennis venom peptides and less than half can be bioinformatically annotated. The paucity of annotation information complicates the design of studies that seek to better understand the potential mechanisms underlying the envenomation response. Although the RNA interference system of N. vitripennis provides an opportunity to functionally characterise venom encoding genes, with 79 candidates this represents a daunting task. For this reason we were interested in determining the expression levels of venom encoding genes in the venom gland, such that this information could be used to rank candidate venoms. To do this we carried out deep sequencing of the transcriptome of the venom gland and neighbouring ovary tissue and used RNA-seq to measure expression from the 79 venom encoding genes. The generation of a specific venom gland transcriptome dataset also provides further opportunities to investigate novel features of this highly specialised organ. Results High throughput sequencing and RNA-seq revealed that the highest expressed venom encoding gene in the venom gland was a serine protease called Nasvi2EG007167, which has previously been implicated in the apoptotic activity of N. vitripennis venom. As expected the RNA-seq confirmed that the N. vitripennis venom encoding genes are almost exclusively expressed in the venom gland relative to the neighbouring ovary tissue. Novel peptides appear to perform key roles in N. vitripennis venom function as only four of the highest 15 expressed venom encoding genes are bioinformatically annotationed. The high throughput sequencing data also provided evidence for the existence of an additional 471 novel genes in the Nasonia genome that are expressed in the venom gland and ovary. Finally, metagenomic analysis of venom gland transcripts identified viral transcripts that may play an important part in the N. vitripennis venom function. Conclusions The expression level information provided here for the 79 venom encoding genes provides an unbiased dataset that can be used by the N. vitripennis community to identify high value candidates for further functional characterisation. These candidates represent bioactive peptides that have value in drug development pipelines.
Project description:Callobius koreanus (C.koreanus) is a wandering spider and a member of the Amaurobiidae family, infraorder Araneae. RNA-sequencing was performend for venom gland tissue and whole body except venom gland.
Project description:Agelena koreana is indigenous spider in South Korea that lives on piles of trees building webs. RNA-sequencing was performed for venom gland tissue and whole body except venom gland.
Project description:We generated ATAC-seq data for pre- and post-extraction venom gland samples and H3K4me3, H3K27ac, and CTCF ChIP-seq from post-extraction venom gland samples from the Prairie Rattlesnake to investigate patterns of chromatin accessibility, transcription factor binding, and insulation during venom production, and to identify open promoters and active enhancer regions.
Project description:The nose-horned viper, its nominotypical subspecies Vipera ammodytes ammodytes (Vaa) in particular is, medically, the most relevant snake in Europe. The local and systemic clinical manifestations of poisoning by the venom of this snake are the result of the pathophysiological effects inflicted by enzymatic and non-enzymatic venom components acting, most prominently, on blood, cardiovascular and nerve systems. This venom comprises the most complex mixture of pharmacologically active proteins and peptides of all European snakes. To help improve the current antivenom therapy towards higher specificity and efficiency, and to assist drug discovery, we have constructed, by combining transcriptomic and proteomic analyses, the most comprehensive library yet of the Vaa venom proteins and peptides. At the protein level, 57 venom proteins belonging to 16 different protein families have been identified and, with SVSPs, sPLA2s, snaclecs and SVMPs, comprise about 80% of all venom proteins.
Project description:Towards a better understanding of molecular processes underlying the acquisition of immune-tolerance during VIT (venom immunotherapy), we analysed genome-wide gene expression profiles of patients with severe hymenoptera venom allergy (HVA), who were analyzed prior to VIT and 12 months after the beginning of a VIT. The study also includes individuals withhout signs of HVA (as revealed by skin tests); beekepers and HVA patients sampled between 1st and 2nd month of VIT.