Search for: All records

Abstract CRISPR-Cas mediated genome engineering has revolutionized functional genomics. However, understanding of DNA repair following Cas-mediated DNA cleavage remains incomplete. Using Cas12a ribonucleoprotein genome editing in the fungal pathogen, Magnaporthe oryzae , we detail non-canonical DNA repair outcomes from hundreds of transformants. Sanger and nanopore sequencing analysis reveals significant variation in DNA repair profiles, ranging from small INDELs to kilobase size deletions and insertions. Furthermore, we find the frequency of DNA repair outcomes varies between loci. The results are not specific to the Cas-nuclease or selection procedure. Through Ku80 deletion analysis, a key protein required for canonical non-homologous end joining, we demonstrate activity of an alternative end joining mechanism that creates larger DNA deletions, and uses longer microhomology compared to C-NHEJ. Together, our results suggest preferential DNA repair pathway activity in the genome that can create different mutation profiles following repair, which could create biased genome variation and impact genome engineering and genome evolution. 

Abstract Located on the campus of the Thacher School in Southern California, the Thacher Observatory has a legacy of astronomy research and education that dates back to the late 1950s. In 2016, the observatory was fully renovated with upgrades including a new 0.7 m telescope, a research grade camera, and a slit dome with full automation capabilities. The low-elevation site is bordered by the Los Padres National Forest and therefore affords dark to very dark skies allowing for accurate and precise photometric observations. We present a characterization of the site including sky brightness, weather, and seeing, and we demonstrate the on-sky performance of the facility. Our primary research programs are based around our multi-band photometric capabilities and include photometric monitoring of variable sources, a nearby supernova search and followup program, a quick response transient followup effort, and exoplanet and eclipsing binary light curves. Select results from these programs are included in this work which highlight the broad range of science available to an automated observatory with a moderately sized telescope.