Intracellular Delivery of mRNA for Cell‐Selective CRISPR/Cas9 Genome Editing using Lipid Nanoparticles

Abstract

Messenger RNA (mRNA) is being used as part of an emerging class of biotherapeutics with great promise for preventing and treating a wide range of diseases, as well as encoding programmable nucleases for genome editing. However, mRNA's low stability and immunogenicity, as well as the impermeability of the cell membrane to mRNA greatly limit mRNA's potential for therapeutic use. Lipid nanoparticles (LNPs) are currently one of the most extensively studied nanocarriers for mRNA delivery and have recently been clinically approved for developing mRNA-based vaccines to prevent COVID-19. In this review, we summarize the latest advances in designing ionizable lipids and formulating LNPs for intracellular and tissue-targeted mRNA delivery. Furthermore, we discuss the progress of intracellular mRNA delivery for spatiotemporally controlled CRISPR/Cas9 genome editing by using LNPs. Finally, we provide a perspective on the future of LNP-based mRNA delivery for CRISPR/Cas9 genome editing and the treatment of genetic disorders.

Unleashing the Power of Proenzyme Delivery for Targeted Therapeutic Applications Using Biodegradable Lipid Nanoparticles

“…The strategic modification of ionizable lipid molecular structures profoundly influences the encapsulation efficiency, intracellular delivery efficacy, and cell-targeting precision of proteins and nucleic acids. The pivotal nature of these lipids necessitates the discovery of novel lipid or lipid-like materials to optimize protein delivery . Empirical evidence from recent investigations has illuminated that through the molecular engineering of ionizable lipids, it is feasible to direct the delivery of mRNA to specific organs such as the liver, spleen, and lung .…”

“…The pivotal nature of these lipids necessitates the discovery of novel lipid or lipid-like materials to optimize protein delivery. 28 Empirical evidence from recent investigations has illuminated that through the molecular engineering of ionizable lipids, it is feasible to direct the delivery of mRNA to specific organs such as the liver, 29 spleen, 30 and lung. 31 However, the incomplete comprehension of the structure−activity dynamics that underpin lipid-mediated protein delivery necessitates the employment of highthroughput screening methodologies.…”