Project description:we used next-generation sequencing technology to characterise mRNA-seq of brackish water (BW, 10‰), fresh water (FW, 0‰), and sea water (SW, 25‰)-treated Anguilla marmorata's gill, kidney and intestine to elucidate the molecular mechanisms of salinity adaptation.
Project description:Aedes aegypti, the principle global vector of arboviral diseases, has been widely regarded to only lay eggs and undergo preimaginal development in fresh water collections. Recent observations however show that it has adapted to develop in anthropogenic brackish water habitats of up to 50% sea water in coastal areas in different continents. This adaptation is characterised by greater salinity tolerance in adult oviposition preference, larvae and changes in sizes of anal papillae. The physiological basis for salinity tolerance in either Ae. aegypti or any of the known salinity-tolerant species of Anopheles malaria vectors is not established. To address this knowledge gap which is of fundamental biological interest and important for control of major diseases we performed RNAseq analysis of gut, anal papillae, and rest of the carcass of Ae. aegypti collected in the field from brackish water (BW) and fresh water habitats (FW) and then maintained as laboratory colonies in BW and FW respectively. We also examined the cuticle structure of larvae, pupae and adult BW and FW Ae. aegypti by microscopy and performed proteomic analysis of the shed cuticles of fourth instar larvae (L4) when they transformed into pupae. The results show that major changes in cuticle structure and composition characterize, and may be the principal factor that permits, the adaptation of Ae. aegypti to brackish water.
Project description:Forward genetic screens across hundreds of diverse cancer cell lines have started to define the genetic dependencies of proliferating human cells and how these vary by genotype and lineage. Most screens, however, have been carried out in culture media that poorly resemble metabolite availability in human blood. To explore how medium composition influences gene essentiality, we performed CRISPR-based screens of human cancer cell lines cultured in traditional versus human plasma-like medium (HPLM). Sets of medium-dependent fitness genes span several cellular processes and can vary with both natural cell-intrinsic diversity and the specific combination of basal and serum components that comprise typical culture media. Notably, we traced the causes for each of three conditional CRISPR phenotypes to the availability of metabolites uniquely defined in HPLM versus traditional media. Our findings reveal the profound impact of medium composition on gene essentiality in human cells, and also suggest general strategies for using genetic screens in HPLM to uncover new cancer vulnerabilities and gene-nutrient interactions.
