Non-viral delivery of CRISPR/Cas9 DNA for gene editing via multivalent cationic liposome system

Abstract

[Excerpt] Introduction: CRISPR/Cas9 gene editing technology has revolutionized medical research by opening new therapeutic possibilities for disease treatment such as cancer, cardiovascular, neuronal and immune disorders [1]. Even with the emergence of this technology, the lack of clinically viable delivery systems continues to hinder CRISPR therapeutic applications. Viral vectors have been sucessfully used for the delivery of CRISPR/Cas9, which whilst efficient, raise safety concerns regarding immunogenicity and insertional mutagenesis [2]. Non-viral vector such as cationic liposomes offer an attractive alternative to viruses given their low immunogenicity, high encapsulation capacity, easy synthesis and functionalization [3]. Their cationic charge mediates strong electrostatic interactions with the negative charges of nucleic acids, leading to the formation of complexes called “lipoplexes”. However, cationic lipids efficiency is still not at the level of viruses as delivery vectors. In recent years, multivalent cationic lipids have been proposed as a promising strategy to effectively deliver nucleic acids into target cells [4]. In this work, we explored the potential of multivalent cationic liposomes to deliver plasmid-based CRISPR/Cas9 systems as well as to mediate gene-editing. [...]