Advancing Gene Therapies through Targeted mRNA Delivery and Scalable CRISPR Solutions

Summary

The presentation by Shambavei and Jian Pang from Jensen Script focuses on advancements in mRNA production, lipid nanoparticles (LNPs) for gene editing, and CRISPR-based therapeutics. They discuss optimizing mRNA, enhancing LNP delivery systems, and improving gene-editing efficiency, particularly for gene knockouts, knock-ins, and prime editing. The presentation also covers the regulatory landscape, risk assessment techniques, and how Jensen Script ensures safety and efficacy through NGS testing and GMP compliance. The company’s comprehensive services aim to advance gene-editing therapies while addressing off-target effects and regulatory requirements.

Takeaways

• 😀 Jensen Script has been in the life sciences industry for nearly 20 years, specializing in mRNA and LNP (lipid nanoparticle) technologies for gene editing solutions.
• 😀 Jensen Script offers three types of mRNA: linear, circular, and self-amplifying, alongside proprietary LNPs to facilitate mRNA delivery to specific tissues or cells.
• 😀 One of the main challenges in gene editing is optimizing LNP formulations, and Jensen Script provides targeted LNPs to enhance tissue-specific delivery and gene editing efficiency.
• 😀 mRNA-based gene editing using CRISPR, specifically using LNPs and guide RNAs, avoids the limitations of viral vectors, such as immunogenicity and cargo size restrictions.
• 😀 Jensen Script uses AI-driven tools for optimizing mRNA codons to increase expression levels, which is critical for gene editing applications like CRISPR.
• 😀 The company has had success with gene knockout and knock-in experiments, including using short and long DNA donor sequences encapsulated in LNPs, achieving greater than 60% knock-in efficiency.
• 😀 The optimization of LNP formulations is key for improving CRISPR editing efficiency, especially when targeting specific genes like PCSK9 for gene therapy applications.
• 😀 Jensen Script's mRNA and LNP technologies have also been adapted for base editing and prime editing, which are cutting-edge gene editing techniques that don't involve double-stranded DNA breaks.
• 😀 Prime editing, a more precise editing method, requires long guide RNAs (around 200 nucleotides) and optimized mRNA-LNP formulations for higher editing efficiency, especially in liver cells.
• 😀 Jensen Script also focuses on improving guide RNA synthesis and ensuring high purity and sterility for regulatory compliance, including through the use of NGS (Next Generation Sequencing) to detect impurities and off-target mutations.
• 😀 Regulatory hurdles for gene editing products are significant, with FDA and EMA requiring precise documentation of purity, sequence identity, and impurity testing through NGS, to ensure safety for clinical use.

Q & A

What is the primary focus of the presentation by Shambavei and Jian Pang?

The presentation focuses on advancements in gene editing using mRNA and lipid nanoparticles (LNPs), specifically in the context of improving mRNA quality, optimizing delivery systems, and supporting gene editing techniques like CRISPR and prime editing.

How does GenScript contribute to mRNA production and gene editing research?

GenScript provides comprehensive solutions in mRNA production, including linear, circular, and self-amplifying mRNA, as well as LNP products for targeted delivery. They also support R&D in mRNA design and optimization, helping advance gene editing technologies.

Why are LNPs preferred over viral vectors for mRNA delivery?

LNPs are preferred because they have no size limitations for cargo, can be redosed, and have lower immunogenicity compared to viral vectors. The toxicity of LNPs can also be minimized through process optimizations.

What are the challenges associated with developing targeted LNPs for gene editing?

Developing targeted LNPs is complex due to the need for multiple lipids, conjugation processes, and antibody tagging. Ensuring that the LNPs effectively target specific tissues or organs requires precise optimization and robust analytical steps.

What role does UTR optimization play in gene editing mRNA design?

UTR optimization enhances the expression of the mRNA, which is critical for efficient gene editing. By screening different UTRs, GenScript was able to identify UTRs that improve expression in liver and other organs, supporting better gene delivery and editing efficiency.

How does GenScript's AI tool assist in gene editing research?

GenScript’s AI tool helps optimize mRNA codons, enhancing mRNA expression. This tool has been used to improve the efficiency of gene editing, such as for prime editing, by boosting expression and increasing editing percentages in cells.

What was the outcome of GenScript’s research on knockout and knock-in gene editing?

GenScript successfully generated knockout and knock-in genes in cells, testing both short and long DNA donor encapsulations. Co-delivery of components significantly improved knock-in efficiency, achieving over 60% editing efficiency with LNPs.

What is the significance of optimizing LNP compositions in gene editing?

Optimizing LNP compositions is crucial for enhancing gene editing efficiency. By adjusting lipid ratios and utilizing proprietary formulations, GenScript was able to improve editing efficiency, particularly for liver cells and genes like PCSK9.

How does GenScript support the development of prime editing systems?

GenScript has developed specialized LNP formulations for prime editing, optimizing the delivery of prime editing enzymes and guide RNAs. These formulations have shown high editing efficiency in various cell lines, including hepatocytes, and are designed for targeted editing in specific cells.

What are the regulatory challenges associated with CRISPR-based gene editing, as mentioned in the presentation?

Regulatory challenges include ensuring guide RNA purity, sequencing identity tests, off-target risk evaluations, and meeting FDA and EMA requirements for clinical trials. GenScript addresses these challenges by streamlining GMP manufacturing and providing necessary documentation and support for regulatory submissions.