Cellular delivery of CRISPR/Cas9 plasmid using multivalent cationic liposomes

Abstract

[Excerpt] Background: Gene therapy techniques have been aimed at mitigating disease features of recessive and dominant disorders, as well as several cancers, cardiovascular, neuronal and immune diseases [1]. In recent years, the clustered regularly interspaced short palindromic repeats (CRISPR) and Cas9 protein system has been gaining recognition as a revolutionary tool for gene therapy. However, the implementation of CRISPR/Cas9 in the clinical setting remains difficult due to challenges in delivery high molecular weight DNA into cells. Various delivery strategies of nucleic acid-based therapeutics, including both viral and nonviral approaches have been tested for the delivery of CRISPR/Cas9 system. Although, the available delivery systems suffer from various limitations, such as immunogenicity and insertional mutagenesis for viral systems [2], and low transfection efficiency for nonviral systems [3]. Multivalent cationic lipids have been proposed as a promising nonviral vector strategy to effectively deliver nucleic acids into target cells. They are characterized by their higher membrane charge density, which has been suggested to facilitate endosomal escape, improving the transfection efficiency [4,5]. In this work, we explored the potential of multivalent cationic liposomes to deliver plasmid based CRISPR/Cas9 systems. [...]