Dalevyon
Knight
104 Program for Research in Science and Engineering Designing and Optimizing Zeocin-Resistant Lentivirus epegRNA and ngRNA Plasmids for Efficient Prime Editing
Abstract profile. Full document pending author claim.
Authors:
Dalevyon Knight, Marc Zepeda, Brian Liau
Date Created:
2025-01-01
Course Title:
Professor:
Not specified
About Paper:
Prime editing is a CRISPR-Cas9-based gene editing technique that additional selectable markers. To address this problem, we introduces precise DNA modifications without double-stranded constructed zeocin-resistant epegRNA and ngRNA vectors using breaks, offering greater accuracy than base editing. In the currenta three-part Gibson assembly. Following bacterial transformation, literature, mammalian prime editing experiments typically require nanopore sequencing, and Geneious software data analysis, our cloning two plasmid vectors: an engineered prime editing guide assemblies showed a 100% match between the synthesized (epegRNA) to provide a template for the reverse transcriptase plasmids and their reference designs, confirming the accuracy and to synthesize the desired edit and a nicking guide (ngRNA) to robustness of this strategy. Overall, our approach may offer a induce a single-strand break in the target DNA. However, current streamlined and efficient strategy for therapeutic plasmid delivery dual-vector systems are limited by inefficient selection in certainin cancer cell lines and could optimize cloning protocols within the cell lines due to Cas9-induced blasticidin resistance, requiring prime editing space.
Abstract:
Prime editing is a CRISPR-Cas9-based gene editing technique that additional selectable markers. To address this problem, we introduces precise DNA modifications without double-stranded constructed zeocin-resistant epegRNA and ngRNA vectors using breaks, offering greater accuracy than base editing. In the currenta three-part Gibson assembly. Following bacterial transformation, literature, mammalian prime editing experiments typically require nanopore sequencing, and Geneious software data analysis, our cloning two plasmid vectors: an engineered prime editing guide assemblies showed a 100% match between the synthesized (epegRNA) to provide a template for the reverse transcriptase plasmids and their reference designs, confirming the accuracy and to synthesize the desired edit and a nicking guide (ngRNA) to robustness of this strategy. Overall, our approach may offer a induce a single-strand break in the target DNA. However, current streamlined and efficient strategy for therapeutic plasmid delivery dual-vector systems are limited by inefficient selection in certainin cancer cell lines and could optimize cloning protocols within the cell lines due to Cas9-induced blasticidin resistance, requiring prime editing space.
Source:
Harvard / Harvard College | Lowell House | Neuroscience | 2027 / 2025
Topics:
editing, prime, plasmid, epegrna, ngrna, zeocin, efficient, dna, vector, break, accuracy, assembly
