mRNA Applied Vector

Specification

Description:

GenScript’s proprietary 2nd generation mRNA vectors incorporate a T7(AG) promoter and wild-type HBA UTR sequences, enabling direct ORF insertion for efficient in vitro transcription. Modular multiple cloning sites are included to accommodate customized promoter or UTR designs as needed.

Plasmid Type	Cloning/Expression
Promoter	T7/CMV promoter
Resistance	Kanamycin
Poly(A) Tract	80A/100A/120A/30+30+43A/31+71A
Linearization Site	BspQI/BsaI

Supporting Data

Engineered for Superior Poly(A) Stability

pUC57 and GenScript mRNA Applied Vectors (pGS), containing Poly(A) tails of 100A and 120A with a GFP insert, were constructed and subcultured into GenStable™ Poly(A) and NEB Stable strains over eight generations. Sanger sequencing was conducted at each generation to evaluate the integrity of the Poly(A) sequences. The pGS-mRNA vector demonstrated significantly superior performance in maintaining Poly(A) sequence integrity during plasmid preparation.

Engineered for Superior Plasmid Production

Genes with various inserts, including toxic and unstable sequences, were cloned into pUC57, pGS-mRNA first-generation, and second-generation vectors containing a 120A poly(A) tail. Following overnight propagation in 400 mL culture, plasmid yield was extracted and evaluated. The pGS-mRNA second-generation vector demonstrated more than a 100% increase in plasmid yield compared to the commercially available pUC57 vector.

Engineered for High-Purity Monomeric Supercoiled Plasmids

Genes with various inserts, including toxic and unstable sequences, were cloned into pUC57, pGS-mRNA first-generation, and second-generation vectors containing a 120A poly(A) tail. Compared to the commercially available pUC57 vector, the second-generation pGS-mRNA vector demonstrated an average 37% increase in monomeric supercoil content.

FAQ

• 1. How does the second generation mRNA applied vector differ from the first generation?

  While both generations demonstrate the same strong ability to maintain Poly(A) stability, the second-generation vector is further engineered to deliver higher plasmid yield and increased monomeric supercoil percentages. Specifically, it achieves ~50% higher plasmid yield compared to the first generation and ~100% higher yield compared to commercially available vectors. On average, monomeric supercoil content increases by ~20% over the first generation and ~40% compared to commercial vectors.

• 2. Why does supercoiling matter for IVT mRNA template preparation?

  Based on our studies, the isoform percentage of supercoiling does not have a direct impact on yield or IVT reaction efficiency. However, maintaining consistent levels of monomeric supercoil is critical for ensuring batch-to-batch quality control. This consistency is especially important for documentation purposes during regulatory submission and agency approval.

• 3. Are GenScript’s mRNA applied vectors built with T7 and UTR sequences, and can I replace them with my own?

  Yes. GenScript’s second-generation mRNA vectors are preconfigured with a T7 promoter (AG) and wild-type HBA UTR sequences, enabling direct cloning of your gene insert for IVT experiments. If you prefer to use your own promoter or UTR design, the vectors also include flexible cloning sites to accommodate customization.

• 4. What are the terms of use for GenScript’s mRNA applied vectors?

  GenScript’s mRNA applied vectors are provided under a limited, non-transferable right for research use only. No license is required for research purposes. However, if you plan to use the vectors for commercial applications, a license must be obtained from GenScript. For more information: https://www.genscript.com/genscript-proprietary-vector-terms.html

Resources

Application Note:
Overcoming Traditional Limitations: GenScript’s Advanced mRNA Applied Vector for Superior Cloning and Efficient Plasmid Production

In response to the growing demand for mRNA-based therapeutics, we have engineered the GS-mK and GS-CMV vectors to address the limitations of traditional vectors such as pUC57 and pVAX1.

Free Download

Application Note:
GenScript Poly(A) Strain: Overcoming Challenges in Pertaining Long Poly(A) Tail Sequence Integrity

To address the challenges associated with maintaining and prorogating plasmids containing poly(A) sequences, GenScript have developed a suite of exclusive Poly(A) strains.

Free Download

Poster:
Enhancing Poly(A) Tail Stability and Plasmid Yield: GenScript's Innovative Vector and Strain Solutions for mRNA Production

Correlation Between Poly(A) Tail Length and Protein Expression. Methods to Enhance Poly(A) Tail Stability During Plasmid Propagation

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