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Custom IVT Synthesis of mRNA

Background IVT Synthesis Services Highlights FAQs Published Data

Background

Creative Biolabs is a leader in biotechnology, providing Custom IVT Synthesis of mRNA tailored for diverse research and clinical applications. Our team, with over ten years of experience in mRNA synthesis and modifications, delivers IVT services characterized by exceptional purity and biological activity. IVT mRNA, crucial in regenerative medicine, vaccinations, and cell engineering, offers a safe, effective gene therapy vector. We are dedicated to propelling gene therapy innovations and support all scientific endeavors with our expert, reliable solutions.

IVT Synthesis Services

By sending us the DNA plasmid or the gene sequence, Creative Biolabs can synthesize a high-quality mRNA depending on the customers' experimental purposes. IVT RNA synthesis requires DNA templates, enzymes, nucleotides, and buffer components. With the experience of the IVT system, Creative Biolabs can offer optimization of each reaction component and several internal epigenetic modifications to obtain high-yield synthesized IVT mRNA for our customers, including but not limited to:

  • Various grades and types of templates design (plasmid DNA, PCR products, and synthetic oligonucleotides)
  • Selection of the best-fit promoter system (such as T7)
  • Codon optimization
  • Optimization of 5'-UTR and 3'-UTR
  • Optimization of poly(A) length and 5' capping

Aiming for protein expression or translation process through synthesized mRNA, all these options are important for improved stability and translation efficiency. Moreover, we can also offer the modifications of mRNA after synthesis to achieve the specific requirements of our customers. To obtain a pristine IVT mRNA, we also offer the large-scale purification step (using PAGE or RP-HPLC methods) to remove the residual molecules, such as unreacted nucleotides, short oligonucleotides, enzyme proteins, and residual salts. These methods remove most of the unwanted byproducts and provide high-quality IVT mRNAs. Our IVT synthesis strategy is very suitable for particularly long mRNAs (such as those up to multiple kilobases), and our experts can design custom strategies to optimize the yield even for the most complicated custom mRNA production requirements.

In addition, we can also provide self-amplifying RNA (saRNA), which is an advanced form of RNA technology designed to enhance the expression of encoded proteins. Unlike conventional mRNA, saRNA includes additional sequences that allow it to replicate itself within the host cell, thereby amplifying the production of the target protein. This technology has significant implications for vaccine development, gene therapy, and protein production.

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Highlights

  • Customized mRNA from 1000 to several thousands of bases
  • Flexible choice of approaches depending on the use of mRNA
  • Numerous strategies for capping and introduction of poly(A) tails
  • Well-characterized mRNA synthesis and purification processes
  • Feasibility of scaling-up to industrial-scale manufact
  • Sterility and bioburden controls
  • Readiness for most downstream applications
  • High-quality services with competitive prices

FAQs

Q: What is the turnaround time for IVT mRNA synthesis?

A: The turnaround time varies depending on the complexity and scale, but Creative Biolabs aims for a fast and efficient process, typically within a few weeks.

Q: What are the typical applications of IVT mRNA?

A: Applications include therapeutic development, vaccine production, functional genomics, and cell reprogramming.

Q: How does Creative Biolabs ensure the quality of synthesized mRNA?

A: We employ rigorous quality control measures including purity checks, integrity analysis, and functionality assays.

Q: What types of promoters are used in IVT synthesis?

A: They use various promoters like T7, SP6, and T3, depending on the specific requirements of the project.

Q: Can Creative Biolabs synthesize modified nucleotides in IVT mRNA?

A: Yes, we offer incorporation of modified nucleotides to enhance mRNA stability and reduce immunogenicity.

Q: What customization options are available in Creative Biolabs?

A: Customization includes various DNA template designs, promoter systems, codon optimization, 5'- and 3'-UTR optimization, and poly(A) tail length adjustments.

Published Data

This article focuses on improving the synthesis of high-integrity mRNA via in vitro transcription (IVT). The main challenge addressed is the generation of fragmented mRNA during the synthesis process, which impedes the efficacy of mRNA therapeutics. The study identifies specific domains in T7 RNA polymerase responsible for premature termination and mRNA fragmentation. By engineering T7 RNA polymerase mutants and optimizing IVT parameters, the researchers achieved mRNA integrity exceeding 91%. The research emphasizes the importance of full-length mRNA transcripts for effective in vivo expression and highlights strategies to reduce impurities, thereby enhancing the potential of mRNA-based therapies.

Capillary electrophoresis results of mRNA produced by in vitro transcription.

Fig.1 Analysis of mRNA products by capillary electrophoresis method.1

Featured mRNA Products

Hot IVT Vectors

Cat. No Product Name Promoter
CAT#: GTVCR-WQ001MR IVTScrip™ pT7-mRNA-EGFP Vector T7
CAT#: GTVCR-WQ002MR IVTScrip™ pT7-VEE-mRNA-EGFP Vector T7
CAT#: GTVCR-WQ003MR IVTScrip™ pT7-VEE-mRNA-FLuc Vector T7
CAT#: GTVCR-WQ87MR IVTScrip™ pT7-VEE-mRNA-Anti-SELP, 42-89-glycoprotein Vector T7

Hot IVTScrip™ mRNA Transcript

Cat. No Product Name Type
CAT#: GTTS-WQ001MR) IVTScrip™ mRNA-EGFP (Cap 1, 30 nt-poly(A)) Reporter Gene
CAT#: GTTS-WK18036MR IVTScrip™ mRNA-Human AIMP2, (Cap 1, Pseudo-UTP, 120 nt-poly(A)) Enzyme mRNA
(CAT#: GTTS-WQ004MR) IVTScrip™ mRNA-Fluc (Cap 1, 30 nt-poly(A)) Reporter Gene
(CAT#: GTTS-WQ009MR) IVTScrip™ mRNA-β gal (Cap 1, 30 nt-poly(A)) Reporter Gene

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Reference

  1. He, Wei, et al. "Effective Synthesis of High-Integrity mRNA Using In Vitro Transcription." Molecules 29.11 (2024): 2461. Distributed under Open Access license CC BY 4.0, without modification.
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