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:Bites by the Indian spectacled cobra (Naja naja) are widely reported across the Indian subcontinent, with an associated high rate of mortality and morbidity. In western India (WI), the numbers of reported incidents of cobra envenomation are significantly higher than the other snake bites. In this study the venom proteome of WINn was deciphered for the first time using tandem mass spectroscopy analysis.
Project description:Latest advancement of omics technologies allows in-depth characterization of venom compositions. In the present work we present a proteomic study of two snake venoms of the genus Naja i.e. Naja naja (black cobra) and Naja oxiana (brown cobra), of Pakistani origin. The present study has shown that these snake venoms consist of a highly diversified proteome. Furthermore, the data also revealed variation among closely related species. High throughput mass spectrometric analysis of the venom proteome allowed to identify for the N. naja venom 34 protein families and for the N. oxiana 24 protein families. The comparative evaluation of the two venoms showed that N. naja consists of a more complex venom proteome than N. oxiana venom.
Project description:Snakebite affects ~1.8 million people annually. The current standard of care are antibody-based antivenoms, which are difficult to access and are not effective against local tissue injury, the primary cause of morbidity. Here we use a functional genomics approach to define human genes that genetically interact with spitting cobra venoms. Most genes that confer resistance to venom cytotoxicity control proteoglycan biosynthesis, suggesting heparinoids as possible inhibitors. Here we show that heparinoids prevent venom cytotoxicity by inhibiting three-finger cytotoxins. Critically, the FDA-approved heparinoid tinzaparin was found to reduce tissue damage in vivo when given via a medically relevant route and dose. Overall, our systematic molecular dissection of cobra venom mechanisms provides new insight into how we can treat cobra bites, information that can help improve the lives of millions of people worldwide.
Project description:This project mainly aims to characterize the complex toxic components present in the venom of Indian cobra (Naja naja) from the Western Ghats of India. Naja naja (NN) is native to the Indian subcontinent and is also found in Pakistan, Sri Lanka, Bangladesh and Southern Nepal. It is a highly venomous snake species of genus Naja of the Elapidae family. They are seen in wide habitats like plains, dense or open forests, rocky terrains, wetlands, agricultural lands, and outskirts of villages and even in highly populated urban areas. This species has been included in the ‘Big 4’ category of venomous snake species that accounts for majority of morbidity and mortality cases in India. Therefore, exploring the venom proteome of Naja naja is decisive to develop and design new antivenom and therapeutics against its envenomation. The venom proteome of Naja naja was characterized through various orthogonal separation strategies and identification strategies. In order to achieve this the crude venom components were resolved on a 12% SDS page. Also, the venom was decomplexed through reversed-phase HPLC followed by SDS analysis. Further each of the bands were subjected to in-gel digestion using trypsin, chymotrypsin and V8 proteases. All the digested peptides were then subjected to Q-TOF LC-MS/MS analysis.
Project description:The synthesis of snake venom proteins is subjected to finely regulated processes in the specialized secretory epithelium within the venom gland. Such processes occur within a defined time frame in the cell and at specific cellular locations. Thus, the determination of subcellular proteomes allows the characterization of protein groups for which the site may be relevant to their biological roles, thereby allowing the deconvolution of complex biological circuits into functional information. In the case of snake venom glands, subcellular proteome analysis could help understand the molecular basis of venom variability. Consequently, knowing the functional implications of such phenotypic plasticity could prove relevant in envenoming treatment and biological research. In this regard, we performed subcellular fractionation of proteins from B. jararaca snake venom gland, focusing on nuclear proteins since this cellular compartment comprises key effectors that shape gene expression. Our results provided a snapshot of B. jararaca's subcellular venom gland proteome. They pointed to a 'conserved' proteome core among different life stages (newborn and adult) and between genders (adult male and female).
Project description:Snake venom gland cDNA sequencing using the Oxford Nanopore MinION with tissue derived from adult venom glands of the painted saw-scaled viper, Echis coloratus
Project description:Complementary bottom-up MS/MS analyses contributed to complete a locus-resolved venom phenotype map for O. hannah, the world's longest venomous snake and a species of medical concern across its wide distribution range in forests from India through Southeast Asia. Its venom composition convincingly explains the main neurotoxic effects of human envenoming caused by king cobra bite. The integration of efficient chromatographic separation of the venom components, and locus-resolved toxin identification through top-down and bottom-up MS/MS-based species-specific database searching, promises a bright future to the field of venom research.