NSF Award
2220587
Cephalopods offer tremendous opportunities for biological discovery. Their novelties include the most complex invertebrate nervous systems and behaviors, the ability to create near-perfect camouflage, unique body plans, taxon specific genes and gene family expansions, specialized organs for hosting microbes, and an enormous capacity to edit genetic information within mRNA. There is a large and diverse cephalopod research community that takes advantage of these unique features for neuroscience, developmental biology, physiology, biomechanics, microbial-host interactions, material properties, genomics, and molecular biology. Despite these biological attributes, cephalopod research has been limited because specimens are difficult to obtain and there is a complete lack of genetically tractable model species. To this end, we are developing gene manipulation and husbandry methods and resources for Euprymna berryi, the hummingbird bobtail squid, a species native to Japan that has proven to be robust in laboratory settings. As part of this effort, we will develop the technologies to drive reporter gene expression and use these methods to create transgenic lines and resources to serve the research community. The tools and methods developed in this work will open up new areas of investigation for the large and diverse community that studies cephalopod biology, and we will hold laboratory-based hands-on workshops to rapidly share our technical advances. Through this program, we have also created an internship program to provide mentorship, professional development, and on-the-job training to aid upper-level students pursuing careers as aquarists and researchers using aquatic organisms.<br/> <br/> A key bottleneck in the study of cephalopod biology is the absence of a genetically tractable model species. This is, in part, due to the relative difficulty of culturing cephalopods in laboratory settings, the difficulty of accessing and manipulating early embryonic stages, and the absence of molecular and sequencing resources for this taxon. To address this these limitations, our goal is to develop the methods to create transgenic lines of Euprymna berryi, a culturable bobtail squid, and then to use these methods to create resources to serve the research community. As part of this effort, we will optimize protocols for creating transgenics by testing random and site-specific insertion of DNA cassettes. We will also create sequence resources for the identification of promoters and enhancers to drive tissue-specific and ubiquitous reporter gene expression. Finally, we will establish robust husbandry conditions for maintaining these lines across multiple generations. This work is expected to have a strong impact on the cephalopod research community by providing new tools and techniques that will open novel avenues of inquiry, making cephalopod biology accessible to both new and established research labs.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
