Sandia Labs FY21 LDRD Annual Report


Engineering the future of renewable biofuels/bioproducts through phycoviruses. Domestic production of next generation renewable biofuels and bioproducts would enhance U.S. energy security. The team discovered that by genetically engineering a new class of viral vectors based on the ubiquitous class of viruses that naturally infect algae, a new system, capable of introducing entire metabolic pathways into algae, could be facilitated. The genetically tractable algal species Tetraselmis striata, known to be easily infected with an isolated virus, is emerging as a potential biotechnology strain and selected as the project’s algal/viral model system. A novel DNA virus, TsV-N1, that infects T. striata has been isolated with a much smaller genome (30 kB) than most algal viruses, so the team engineered a specialized transducing virus from the TsV-N1 viral chassis to transfer specific, defined DNA fragments with the virus to the host upon infection. Georgia Tech, one of Sandia’s academic partners, was instrumental in identifying a potential chemical compound that can protect algae from deleterious species and prevent pond crashes. These compounds represent several “lead targets” for the engineering of resistance into microalgal production strains through the types of mechanisms that Sandia is developing. Progress made in the first year of this three-year project already represents a significant advance in the ability to rapidly create wholescale genetic changes in microalgae—an important first step toward production of critical renewable biofuels. (PI: Todd Lane) New antibody capable of detecting genome editors. CRISPR-based genome editing , most commonly using Cas9, revolutionized biomedical research and is poised to revolutionize medical treatment, with human clinical trials having begun in 2016. Current tools available for detection of Cas9 focus on bulk detection. No current technology can rapidly detect Cas9 expression within individual live cells. This LDRD project harnessed the naturally occurring

antigen-presenting properties of mammalian cells to develop tools to detect intracellular Cas9 and resulted in a detailed understanding of how CRISPR is delivered to and used in humans, both experimentally and in silico methods to understand and predict the cellular immune response to Cas9, and the development of the first antibody capable of detecting immune presentation of Cas9 peptides or other genome editors. The tools developed through this project enable (1) rapid prediction and understanding of immune response to foreign proteins and (2) the development of detection reagents for foreign intracellular proteins that apply broadly to understanding and enhancing genome editing and combating intracellular infections. (PI: Kimberly Butler)

Method to detect intracellular Cas9 utilizing the native mammalian cell antigen presentation.



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