Research Spotlight: CRISPR Gene Therapy: Applications, Implications, and Avanti!

Posted on January 12, 2022

Gene Editng Res Spot Square For Website

Lipids aren’t just for delivering vaccines

CRISPR/Cas9 is a simple, two-component system that is faster, cheaper, more efficient, and more accurate than its genome editing predecessors. It allows genetic material to be added, removed, or altered at targeted locations in the genome. This ability to change an organism’s DNA represents a promising method for the prevention and treatment of human diseases. As an overview, CRISPR/Cas9 works in two parts: First, by cleaving the DNA strand non-complementary to the spacer sequence thereby generating double-stranded breaks in the target DNA, and (2) by using a single guided RNA (sgRNA) carrying sequence to guide the CRISPR/Cas9 complex to its intended target location. Once the CRISPR/Cas9 complex is in place, one of two endogenous DNA repair pathways are used to edit the DNA sequence. One repair pathway, the non-homologous end-joining (NHEJ) pathway, can result in frameshift deletions because of the random nature of the pathway. The other pathway, the homology-directed repair (HDR) pathway is of more interest in clinical research. The HDR pathway is an error-free repair pathway and uses a template to correctly repair damaged DNA. To take advantage of the HDR pathway, an exogenous template must be used with the CRISR/Cas9 complex to correct the damaged DNA in the desired manner (Sen, et. al, 2020).

So, now that you have a little background information regarding CRISPR/Cas9 genome editing, let’s get to the really interesting part: how Avanti products are being used to aid this gene editing technique!

Using Lipid Nanoparticles (LNPs) to Assist the Hepatic Delivery of Gene Editing Elements for Oncotherapy

Entrance into the target cell is required for therapeutic effectiveness in vaccines. CRISPR/Cas9 is no different in this regard. The complex will not have any effect if it is not properly guided into the target cell. That’s where lipids come into play. Lipids aren’t the only delivery mechanism for gene editing complexes, but they do represent a non-viral, safe, and efficacious delivery tool for long-term administration. With the recent approval of Onpattro®, an siRNA-loaded LNP formulation, and the three recently approved mRNA-based COVID vaccines, LNPs are gaining traction and trust as potent delivery systems for gene editing technology as well.

Ionizable cationic lipids can be used to facilitate membrane fusion, membrane disruption, and endosomal escape due to its ability to be protonated in the acidic environments. Scientists at the Beijing Institute of Technology were able to prepare a lipid nanoparticle delivery system using a novel ionizable lipid, cholesterol, DMG-PEG2000, and DSPC. This lipid nanoparticle system was evaluated for its ability to deliver anticancer CRISPR/Cas9 gene editing systems both in vitro and in vivo.

The study proved that the CRISPR/Cas9 loaded lipid nanoparticle was successfully internalized by the hepatoma carcinoma cells. Furthermore, once internalized, long-term gene editing was achieved in both the in vitro and the in vivo experiment models. The newly developed lipid nanoparticle with the novel ionizable lipid exhibited a more robust nucleic acid delivery than common alternatives. This study shows the potential for robust, effective, and safe gene editing-based cancer treatment. To read the full scope of the study, click HERE!

Avanti offers everything you need to prepare a lipid nanoparticle system capable of delivering your gene editing complexes. We’re your one-stop shop for lipid nanoparticle components including neutral lipids, structural lipids, PEGylated lipids, and cationic lipids. This research utilized a novel ionizable lipid, but Avanti offers several types of cationic lipids such as multivalent cationic lipids, ionizable cationic lipids, and fixed cationic lipids. And if it’s not in our catalog, contact our custom synthesis team to discuss opportunities for us to prepare exactly what you’re looking for. Whether it’s drug-delivery or delivery of gene editing technology, you can trust that you’ll get the best service and the highest quality lipids for your research when you choose Avanti. Click the link to check out our lipid nanoparticle technology page! Avanti's LNP Technology

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