mRNA Gene Delivery Solutions

VectorBuilder provides a one-stop solution for the development of mRNA-based therapeutics, such as vaccines, gene editing, chimeric antigen receptor (CAR), and protein expression in cells or embryos. Based on extensive design and production experience, our team can support researchers for in vitro transcription (IVT) vector design, vector cloning, in vitro mRNA synthesis, LNP-mRNA production, and in vitro/in vivo functional testing to accelerate the development of mRNA-based vaccines and gene therapy. For large-scale manufacturing of mRNA therapeutics, check out our CDMO services. For purchasing ready-to-use mRNA products, check out our catalog IVT mRNA and LNP-mRNA.

Click to view 1000+ citations 

Click to view user testimonials 

Service Details

IVT vector design & cloning IVT vector
design & cloning
IVT mRNA & LNP manufacturing IVT mRNA & LNP manufacturing
Quality control (QC) Quality control (QC)
Functional validation Functional validation
icon_IVT_vector_design_cloning IVT vector design & cloning
  • Royalty-free IVT backbone with no IP constraints for commercial use
  • Sequence optimization for achieving high expression
    • 5’ and 3’ UTR
    • Coding sequence
    • Kozak
  • Robust cloning and transcription of 120 nt (and longer) polyA tail for high expression
IVT_mRNA_LNP_manufacturing IVT mRNA & LNP manufacturing

mRNA synthesis and QC > LNP encapsulation > LNP quality control (RiboGreen assay and dynamic light scattering).

Figure 1. Typical workflow of mRNA synthesis and LNP encapsulation.

  • From vector cloning to LNP encapsulation in as fast as 5 weeks
  • Synthesis of mRNA and self-amplifying RNA (saRNA) of up to 10,000 nt from ug to gram scale
  • High capping efficiency (up to 99%) by co-transcriptional or enzymatic capping
  • Modified nucleotides for reducing immune response and achieving robust expression in vivo
    • m1Ψ
    • m5C
    • 5moU
  • Proprietary purification technology to rapidly and efficiently remove impurities include:
    • IVT DNA template
    • Partially transcribed mRNA products
    • Residual protein
    • Double-stranded RNA (dsRNA)
  • High-quality LNP with conventional and custom formulation:
    • High encapsulation efficiency
    • Low polydispersity index (PDI)
    • Improved stability
    • Increased delivery efficiency
    • Antibody-conjugation compatibility
Quality_control_QC Quality control (QC)

VectorBuilder offers an extensive variety of QC methods for IVT mRNA and LNP encapsulation. Default QC items (marked with √) are always performed while optional QC items are performed depending on individual project needs.

IVT mRNA
LNP encapsulation
Attribute QC Assay Research-grade GMP-like
Identity mRNA sequence Sanger sequencing
mRNA length Denaturing agarose gel electrophoresis
Capillary gel electrophoresis (CGE) Optional
General/physical property mRNA concentration UV spectrophotometry
RiboGreen assay Optional
Appearance Visual inspection Optional
Potency Gene expression In vitro translation followed by Western blot Optional Optional
Cell transfection Optional Optional
Safety Sterility Bioburden test Optional
Mycoplasma Culture method Optional
qPCR Optional Optional
Endotoxin Kinetic chromogenic assay (KCA) Optional
Purity mRNA integrity Denaturing agarose gel electrophoresis
Capillary gel electrophoresis (CGE) Optional
A260/280 UV spectrophotometry
Capping efficiency LC-MS Optional
PolyA analysis LC-MS Optional
Residual dsRNA Dot blot assay Optional
Residual plasmid DNA qPCR Optional
Residual protein NanoOrange assay Optional
Residual solvents Gas chromatography Optional Optional
Attribute QC Assay Research-grade GMP-like
Encapsulation efficiency RiboGreen assay
Particle size, PDI Dynamic light scattering (Zetasizer)
Surface charge Dynamic light scattering (Zetasizer)
Functional_validation Functional validation
  • Assess the effects of different sequence optimizations (UTRs, coding sequence, polyA, Kozak, etc.) on GOI expression in parallel, using our high-throughput cloning, synthesis, and testing platforms.
  • Established functional validation platforms for various applications, such as antigen presentation, antibody expression, CAR expression, and CRISPR.
  • Assess LNP-mRNA gene delivery efficacy and safety using animal models including rodents and non-human primates (NHPs).

Technical Information

IVT vector sequence optimization
IVT mRNA synthesis optimization
LNP-mRNA QC data

VectorBuilder has optimized our encapsulation technology to ensure the homogeneity, stability and efficacy of our LNP encapsulated mRNA. 

LNP-mRNA functional validation
Antibody-conjugated LNP-mRNA

Antibody conjugated LNP-mRNA

Figure 15. Anti-CD31 conjugated firefly luciferase (FLuc) LNP-mRNA showed improved luciferase expression in lung. Mice strain: C57BL/6J; mice age: 6-8 weeks; mice gender: female; administration route: tail vein. Negative controls: IgG2a-conjugated FLuc LNP-mRNA and naked FLuc mRNA.

How to Order

FAQ

What are the differences between mRNA caps and capping methods?

Cap 0 refers to N7-methylguanosine (m7G) that is added to the 5’ end of eukaryotic mRNAs via a 5’ to 5’ triphosphate linkage. This modification is added via a series of enzymatic reactions that occur co-transcriptionally and functions to regulate nuclear export, transcript stability, and promotes translation of the mRNA through recognition by eukaryotic translation initiation factor (eIF4E). Cap 1 refers to the addition of a methyl group to the 2’O on the first nucleotide (m7GpppNm) of the transcribed mRNA sequence in addition to the m7G cap. In mammalian cells, cap 1 structure is an important marker for mRNA to be recognized as self and not targeted by innate immunity. Adding cap 1 structure to synthesized mRNA has been demonstrated to enhance mRNA expression in vivo and reduced its immunogenicity.

Capping for in vitro transcribed RNA can occur either co-transcriptionally with cap analogs or post-transcriptionally via enzymatic reactions. We offer both capping methods, and the efficiency of them has been well validated using LC-MS. Depending on the client-preferred capping method, we will choose a compatible backbone for cloning the IVT mRNA vector from the beginning.

Why should I consider incorporating modified nucleotides in mRNA and which ones can be included?

Cells contain cytosolic and endosomal RNA receptors that activate the immune response upon recognition of foreign RNA. Modified nucleotides are commonly found in endogenous cellular RNA. Incorporating certain modified nucleotides in IVT mRNA reduces its immunogenicity, alters secondary structure, and increases translation efficiency and half-life in a sequence-dependent manner. We provide a wide range of modified nucleotides, including the commonly used N1-Methylpseudouridine (m1Ψ) and 5-Methylcytosine (m5C). N1-Methylpsuedouridine and 5-Methylcytosine are naturally occurring nucleotides that were first identified in tRNAs, however, their use in coding mRNAs has only recently been appreciated. These methylated derivatives of uridine and cytosine can replace their canonical nucleotides in mRNA IVT and translation without altering traditional Watson-Crick base pairing. A major advantage to their use in mRNA therapeutics is their ability to alter recognition by RNA immune receptors thus mitigating unwanted immune effects and enhancing transcript stability and translation.