EXPERIMENTAL PERTURBATION OF THE SEA LAMPREY VARIABLE LYMPHOCYTE SYSTEM USING CRISPR/CAS9
Jonathan Rast, Emory University
Jonathan P. Rast1
1Department of Pathology and Laboratory Medicine, Emory University
Lampreys and hagfish comprise about 60 species of jawless vertebrates that collectively represent an anciently derived sister group to the jawed vertebrates. While, the jawed vertebrates use diversified immunoglobulin (Ig) and T cell receptors (TCR) as the basis of their adaptive immune system, the jawless vertebrates instead use somatically diversified leucine rich repeat proteins, the variable lymphocyte receptors (VLRs). Well-formulated comparisons between these systems potentially provide a long-sought window into the deep origins of vertebrate adaptive immunity. Structurally, the VLR proteins are completely unrelated to Ig/TCR, yet they have analogous functional characteristics and are rooted in a cellular system with distinct similarities to jawed vertebrate B and T cells. Since their discovery 15 years ago, much has been learned about the structure, diversity and expression of the VLRs and their response to immune challenge. Still, fundamental questions remain about how the system functions and how it relates to the jawed vertebrate adaptive system. Many of these lines investigation require new experimental strategies to manipulate the system in vivo. As one approach, we have begun CRISPR/Cas9 experiments to probe causal linkage among VLRs and their predicted assembly and regulator factors in the sea lamprey. We are also interested development of the VLR system in the early feeding larva. I will discuss our findings to date and plans for future research.