Sculpting the genome and beyond: novel tools for DNA and RNA targeting

Abstract

This research delves into the utilization of CRISPR-Cas tools for advanced genome editing. It unveils a novel method known as Ligation-Assisted Homologous Recombination (LAHR), which achieves efficient editing and serves as a robust complement to the established Cas9-induced HDR techniques. The Prime Editor (PE) technology is scrutinized, with a focus on the issue of low editing efficiency. To counter this challenge, an improved variant, PE plus (PE+), is introduced, demonstrating a marked increase in editing efficiency. The study further presents a unique cell selection tool using LbuCas13a, which exhibits widespread RNA cleavage in human cells, hinting at its potential use in cancer therapy. The concluding section foresees a transition in genome editing tools from being DNA repair-dependent to -independent, while also addressing pivotal challenges such as off-target editing and the formulation of effective delivery strategies.