mRNAs for Genome Engineering Services

At Seattle Genova we have a broad range of experience in supplying clients with mRNA products at the appropriate quality level for each program stage. Our proprietary mRNA synthesis technology achieves highly efficient and economical co-transcriptional capping, and we offer numerous post-transcriptional modifications, including DNase and phosphatase treatments, enzymatic capping, and polyadenylation. Additionally, you can select from a variety of purification options, including silica gel purification, liquid chromatography isolation, and high-performance liquid chromatography.

Genome editing, or genome engineering, or gene editing, is a kind of genetic engineering in which DNA is inserted, eliminated, modified or replaced in the genome of a living organism.

Based on engineered or bacterial nucleases, the improvement of genome editing technologies has opened up the chance of directly targeting and modifying genomic sequences in nearly all eukaryotic cells. Genome editing has expanded our ability to elucidate the contribution of genetics to disease by facilitating the creation of more valid cellular and animal models of pathological processes and has started to show extraordinary potential in a variety of fields, ranging from basic research to applied biotechnology and biomedical research.

Recent advances of the three major genome editing technologies (ZFNs, TALENs, and CRISPR/Cas9) and examine the applications of their derivative reagents as gene editing tools in several human disorders and potential future therapies, concentrating on eukaryotic cells and animal models. Ultimately, we give the clinical trials applying genome editing platforms for disease treatment and some of the challenges in the implementation of this technology.

In plants, genome editing is an impressive procedure to alter gene functions to produce improved crop varieties. Genome editing is thought to be simple to utilize and has a lower risk of off-target impacts compared to classical mutation breeding. Similarly, genome-editing technology methods can also be applied directly to crops that include complex genomes and/or are not easily bred utilizing traditional methods.

 Presently, highly versatile genome-editing tools for critical and predictable editing of almost any locus in the plant genome make it feasible to broaden the range of application, containing functional genomics research and molecular crop breeding. Vegetables are important nutrient sources for humans and provide vitamins, minerals, and fiber to diets, thereby providing to human health.

Step 1. Plasmid Manufacturing 

◆ mRNA synthesis begins with plasmid design and production. 

◆ Plasmids are generated in bacterial cultures, then harvested and purified.

Step 2. Transcrption 

In-vitro transcription (IVT)

◆ In vitro transcription is a method that facilitates for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of various kilobases in μg to mg quantities. 

◆ It is based on the engineering of a template that contains a bacteriophage promoter sequence (e.g. from the T7 coliphage) upstream of the sequence of interest fulfilled by transcription utilizing the corresponding RNA polymerase.

Step 3. mRNA purification

After certain manufacturing steps it is significant to purify the mRNA. 

◆ mRNA purification eliminates enzymes, remaining nucleotides, plasmid DNA, and defective mRNA. New emerging technologies like Fibro chromatography, currently accessible for mAb purification, are in development for molecules such as DNA plasmids and mRNA.

Step 4. mRNA encapsulation and polishing

◆ The purified mRNA-based therapeutic is formulated in lipid nanoparticles (LNPs) as a drug delivery vehicle.

◆ Core chromatography can be used to further eliminate impurities.

Step 5. QC release and stability testing

Relying on your final mRNA application and clinical stage, the quality control testing requirements may vary.

◆ RNA content by UV-Vis

◆ Purity by IRRP HPLC

◆ Residual DNA by RT-qPCR

◆ Residual protein by MS

◆ Potency by cell-free translation

◆ Endotoxin and residuals measurements

◆ Sequencing

SERVICE HIGHLIGHTS

◆ High quality products and services at competitive prices

◆ Custom tailored assistance to meet specific application or program needs

◆ Vast variety of modification, treatment, and purification options

◆ Accessible custom synthesis up to gram scales of mRNA and long RNA (multiple   kilobases)

◆ In-house plasmid manufacturing optimized for therapeutic mRNA generation

DELIVERABLES

We provide high throughput evaluations along with faster results. In addition to that, we provide many different mRNA formulation services to meet your various end-point in vaccine delivery. These formulations come with specific functionality to improve the efficiency of vaccines in the physiological environment.

References

1.Abdallah N. A., Prakash C. S., McHughen A. G. (2015). Genome editing for crop improvement: challenges and opportunities. GM Crops Food 6, 183–205.

2.Saurabh S (March 2021). "Genome Editing: Revolutionizing the Crop Improvement". Plant Molecular Biology Reporte. 39: 752–772.