In Vitro Transcription Services

VectorBuilder offers in vitro transcription (IVT)-based synthesis services for custom RNAs up to more than 10 kb in length and suitable for a variety of downstream applications such as non-coding RNAs (miRNA, siRNA, lncRNA, etc.), in situ hybridization, in vitro translation, studying RNA-protein interaction, and as templates in reverse-transcription reactions.

Our RNA synthesis process is highly optimized to produce high-quality RNAs utilizing our proprietary in vitro transcription vectors that are used in our vector cloning services. We can also synthesize RNAs from customer-supplied vectors. In addition to mRNAs, we can generate small RNAs for applications requiring short RNAs of defined sequences including gRNAs for CRISPR/Cas9 targeting, siRNA, miRNA, lncRNA, etc. For developing efficient IVT mRNA, we provide a one-stop mRNA Gene Delivery Solutions including mRNA design, comprehensive quality control (QC), and CRO validation.

Types of RNAs offered
  • RNAs of up to >10 kb in length
  • gRNAs for CRISPR applications
  • Other short RNAs of defined sequences (microRNA, siRNA etc.)

Technical Information

RNA production process and QC

For RNA production, we use the in vitro transcription approach which utilizes the T7 promoter upstream of a DNA template sequence of interest to facilitate highly efficient production of RNA by the bacteriophage T7 RNA polymerase (RNAP), under appropriate reaction conditions and in the presence of nucleotide triphosphates. The T7 RNAP is a highly robust enzyme and transcription reactions using this enzyme can produce large amounts of functional RNA within a few hours. Our typical mRNA production workflow starts with designing and synthesis of the template DNA sequence followed by its cloning into an in vitro transcription vector. The plasmid DNA is then purified, validated, and linearized before being subsequently subjected to the in vitro transcription reaction which results in the generation of the desired transcript. The RNA is then purified by silica gel purification which is our default purification process. We can also purify mRNAs by magnetic bead purification upon request.

Figure 1. Typical workflow of RNA synthesis by in vitro transcription.

All RNAs produced by VectorBuilder undergo stringent quality control to ensure that they are free of contaminants and contain the correct sequence exactly as designed. This involves: 1) validation of the in vitro transcription vector containing the cloned DNA template sequence by restriction digestion analysis and Sanger sequencing; 2) final QC of the synthesized RNAs by denaturing PAGE or agarose gel electrophoresis depending on the size of the transcript.

Documents
Brochures & Flyers

How to Order

Order both vector cloning and RNA preparation
Order RNA preparation from your own vector