A new methodology termed selective organ targeting (SORT) was recently developed that enables controllable delivery of nucleic acids to target tissues. SORT lipid nanoparticles (LNPs) involve the inclusion of SORT molecules that accurately tune delivery to the liver, lungs and spleen of mice after intravenous administration. Nanoparticles can be engineered to target specific cells and organs in the body by passive, active and endogenous targeting mechanisms that require distinct design criteria. SORT LNPs are modular and can be prepared using scalable, synthetic chemistry and established engineering formulation methods. This protocol provides detailed procedures, including the synthesis of a representative ionizable cationic lipid, preparation of multiple classes of SORT LNPs by pipette, vortex and microfluidic mixing methods, physical characterization, and in vitro/in vivo mRNA delivery evaluation. Depending on the scale of the experiments, the synthesis of the ionizable lipid requires 4–6 d; LNPs can be formulated within several hours; LNP characterization can be completed in 2–4 h; and in vitro/in vivo evaluation studies require 1–14 d, depending on the design and application. Our strategy offers a versatile and practical method for rationally designing nanoparticles that accurately target specific organs. The SORT LNPs generated as described in this protocol can therefore be applied to multiple classes of LNP systems for therapeutic nucleic acid delivery and facilitate the development of protein replacement and genetic medicines in target tissues. This protocol does not require specific expertise, is modular to various lipids within defined physicochemical classes, and should be accomplishable by researchers from various backgrounds. This protocol details the preparation of selective organ-targeting lipid nanoparticles by several methods, their physical characterization, and in vitro/in vivo mRNA delivery evaluation. 

/pdf/preparation-of-selective-organ-targeting-sort-lipid-25p5dlpt.pdf

Preparation of selective organ-targeting (SORT) lipid nanoparticles (LNPs) using multiple technical methods for tissue-specific mRNA delivery

Hydroxycholesterol substitution in Ionizable lipid nanoparticles for mRNA delivery to T cells.

mRNA vaccines have emerged as a revolutionary tool to generate rapid and precise immune responses against infectious diseases and cancers. Compared with conventional vaccines such as inactivated viruses, viral vectors, protein subunits or DNA-based vaccines, mRNA vaccines stand out owing to multiple advantages, including simplicity of design, fast production, enhanced safety and high efficacy. Nevertheless, efficient and targeted delivery of mRNA molecules remains a significant challenge owing to their inherent instability and susceptibility to degradation. Nanotechnology offers innovative solutions to surmount these obstacles and amplify the potency of mRNA vaccines. This Primer aims to outline a modular approach to developing biomaterials and nanotechnology for mRNA vaccines, with a focus on particle design, formulation evaluation and therapeutic applications. We delve into the underlying mechanisms of nanoparticle-facilitated mRNA protection, cellular uptake, endosomal escape and immune stimulation. We underscore the critical parameters that impact the manufacturing and clinical implementation of nanomaterial-based mRNA vaccines. Finally, we present the current limitations and future perspectives in the advancement of nanotechnology-enhanced mRNA vaccines for broad applications in prophylactic and therapeutic interventions. mRNA vaccines produce rapid and precise immune responses against infectious diseases and cancers. Chen et al. discuss the development of biomaterials and nanotechnology for mRNA vaccines, how these are designed and evaluated and the underlying mechanisms of cellular uptake and immune stimulation. 

Nanotechnology-based mRNA vaccines

Strategies to reduce the risks of mRNA drug and vaccine toxicity.

Abstract Over the past several decades, mRNA vaccines have evolved from a theoretical concept to a clinical reality. These vaccines offer several advantages over traditional vaccine techniques, including their high potency, rapid development, low-cost manufacturing, and safe administration. However, until recently, concerns over the instability and inefficient distribution of mRNA in vivo have limited their utility. Fortunately, recent technological advancements have mostly resolved these concerns, resulting in the development of numerous mRNA vaccination platforms for infectious diseases and various types of cancer. These platforms have shown promising outcomes in both animal models and humans. This study highlights the potential of mRNA vaccines as a promising alternative approach to conventional vaccine techniques and cancer treatment. This review article aims to provide a thorough and detailed examination of mRNA vaccines, including their mechanisms of action and potential applications in cancer immunotherapy. Additionally, the article will analyze the current state of mRNA vaccine technology and highlight future directions for the development and implementation of this promising vaccine platform as a mainstream therapeutic option. The review will also discuss potential challenges and limitations of mRNA vaccines, such as their stability and in vivo distribution, and suggest ways to overcome these issues. By providing a comprehensive overview and critical analysis of mRNA vaccines, this review aims to contribute to the advancement of this innovative approach to cancer treatment. 

https://molecular-cancer.biomedcentral.com/counter/pdf/10.1186/s12943-023-01807-w

The use of RNA-based treatments in the field of cancer immunotherapy

Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery

Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery.

The cover image is based on the Research Article Rational design of anti-inflammatory lipid nanoparticles for mRNA delivery by Michael John Mitchell et al., https://doi.org/10.1002/jbm.a.37356. 

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Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery

Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery.

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