Authors:

Brian Liau

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About Paper:

Prime editing is a CRISPR-Cas9-based gene editing technique that introduces precise DNA modifications without double-stranded breaks, offering greater accuracy than base editing. In the current literature, mammalian prime editing experiments typically require cloning two plasmid vectors: an engineered prime editing guide (epegRNA) to provide a template for the reverse transcriptase to synthesize the desired edit and a nicking guide (ngRNA) to induce a single-strand break in the target DNA. However, current dual-vector systems are limited by inefficient selection in certain cell lines due to Cas9-induced blasticidin resistance, requiring additional selectable markers. To address this problem, we constructed zeocin-resistant epegRNA and ngRNA vectors using a three-part Gibson assembly. Following bacterial transformation, nanopore sequencing, and Geneious software data analysis, our assemblies showed a 100% match between the synthesized plasmids and their reference designs, confirming the accuracy and robustness of this strategy. Overall, our approach may offer a streamlined and efficient strategy for therapeutic plasmid delivery in cancer cell lines and could optimize cloning protocols within the prime editing space. Detection of High-Energy Neutrino from Supernovae with DUNE Shane Komeiji, Alex Wen, Carlos Argüelles-Delgado Harvard College | Quincy House | Physics | 2028 While a core-collapse supernova can produce over 1050 neutrinos and antineutrinos from nuclear processes, which contain nearly all the energy of the supernova, these particles are difficult to detect on Earth. High-energy supernova neutrinos, which are produced through the collision of an exploding supernova with the surrounding material, can be observed with Cherenkov- based detectors of large masses to maximize the number of neutrino interactions, but full reconstruction of these events is difficult. In this contribution, we will demonstrate the effective detectability of high-energy neutrinos from supernovae with the Deep Underground Neutrino Experiment (DUNE), a liquid argon experiment currently being designed and constructed. We first quantified the total number of starting events in DUNE across all flavors of neutrinos and antineutrinos, assuming optimistic detector efficiencies. We performed an analytical calculation using an existing simulation of type IIn and II-P supernovae neutrino yields with varying densities of circumstellar material. Using SIREN (a simulation software designed to inject and weight particle interactions based on detailed detector geometries), we find the number of neutrinos interacting within the instrumented volume for each flavor. Our preliminary results indicate that, given a supernova at a distance of 10 kiloparsecs, DUNE may detect up to 24 starting neutrino and antineutrino events, about 8 of which are of the tau flavor. Since DUNE is a Liquid Argon Time Projection Chamber, reconstruction and separation of the energetic tracks of the tau neutrinos are easier than with large-volume detectors such as IceCube. This may enable more detailed signals from tau neutrinos and contribute to a deeper understanding of the flavor composition of core-collapse supernovae neutrinos. Harvard Summer Undergraduate Research Village Program for Research in Science and Engineering

Source:

Harvard / Chemistry / 2027

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Co-authors:

Brian Liau