Genome editing based on in vitro-assembled ribonucleoproteins in plants

Abstract

Gene editing technology based on the clustered regularly interspaced short palindromic repeats (CRISPR) system has garnered widespread use in plant genomes because of its cost-effectiveness, efficiency, and simplicity. To avoid the integration of foreign genes and any DNA fragments into target cell genomes, researchers have developed a system that introduces in vitro-assembled ribonucleoproteins (RNPs) consisting of guide RNA (gRNA) and Cas protein into target cells, enabling direct genome editing. This system was designed to deliver RNPs through four distinct methods: polyethylene glycol (PEG)-mediated cell transfection, particle bombardment, electroporation, and lipid transfection. In recent years, CRISPR technology has been extensively applied for the genetic modification of plants, providing a strategic response to environmental challenges. Researchers have successfully established RNP genome editing systems in various plant species. Despite some remaining issues, the RNP genome editing system still shows significant promise for future applications in the production of non-genetically modified (non-GM) crops.