Abstract 2523 CRISPR-Cas as a diagnostic tool for field and conservation biology research and education

In recent years CRISPR-based systems have moved beyond simple gene edits utilizing Cas9.Manipulation of Cas9 (i.e.dCas9) and identification of other Cas variants has further expanded researchers and educators can do with CRISPR, such as CRISPR-based molecular diagnostic assays.CRISPR-based diagnostics have the distinct advantage over their laboratory counterparts including increased sensitivity, ease of use, relatively low cost, and need for minimal laboratory space.To this end, expanded and manipulated a CRISPR-based diagnostics system for deployment in the field as a conservation tool.One effect of global climate change is loss of species diversity: either through expansion of animal ranges or the introduction of invasive species.In some cases, this leads to hybridization of closely related species, which in turn has severe impacts on carefully balanced ecosystems.We used a CRISPR-dCas9 diagnostic test that can detect hybridization of two endemic Antillean turtle subspecies (Trachemys stejnegeri and Trachemys decorata) with the invasive red-eared slider (Trachemys scripta) in the Dominican Republic.This assay uses a simple lateral flow strip assay to detect the presence of T. scripta DNA in blood samples by taking advantage of the high specificity of CRISPR and its ability to distinguish between highly similar DNA sequences.This field CRISPR-dCas9 diagnostic test is simple, cost effective and can be deployed at the point of intervention as a conservation tool.Deployment of this dCas9 assay as integrated into a field course with the explicit goal of providing students the opportunity to participate field work and witness the interdisciplinary nature of science where biochemistry and molecular biology are necessary tools to fully understand major scientific issues like global climate change.