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In Vitro Transcription

An important step in mRNA display is to obtain an mRNA display library. The T7 promoter segment facilitates extensive T7 transcription in vitro to convert the genetic repository into an mRNA collection, furnishing templates for subsequent in vitro translation processes. CD BioSciences offers mRNA display-based drug discovery services to support your early-stage drug discovery efforts.

Peptide Selection Using mRNA Display- CD BioSciences Fig 1. General scheme for peptide selection by mRNA display.

Principles of In Vitro Transcription

In vitro transcription is based on a fundamental principle in molecular biology. This process harnesses natural enzymatic mechanisms present in cells to generate RNA molecules outside the native cellular environment. The precise conditions for the transcription reaction depend on the quantity of RNA required for a specific application. T7 RNA polymerase is an RNA polymerase from the T7 phage and is commonly used for in vitro transcription.

DNA library design. Fig 2. In vitro generation of RNA molecule by a phage T7 RNA polymerase. (Perenkov, et al., 2023)

The in vitro transcription reaction is a crucial step in the synthesis of RNA from a DNA template. The following table shows the key components of the in vitro transcription reaction.

Components	Description
DNA Template	The DNA molecule is the template for RNA synthesis. This DNA template can be obtained through vendor or PCR amplification.
RNA Polymerase	A distinct enzyme, commonly T7, T3, or SP6 RNA polymerase, identifies particular promoter sequences on the DNA template and initiates RNA synthesis complementary to the DNA strand.
Ribonucleotides	The building blocks for RNA synthesis are the four ribonucleotide triphosphates (ATP, UTP, CTP, GTP). The RNA polymerase adds these ribonucleotides one at a time.
Transcription Buffer	A buffered solution provides the optimal conditions for RNA polymerase activity, including pH, ionic strength, and cofactors.

Our Services

Our mRNA display experiments are consistently conducted in vitro, encompassing both the transcription of DNA libraries and the subsequent translation of mRNA libraries. Our transcription methodology for DNA libraries is well-established, enabling us to furnish tailored transcription systems for clients, comprising the necessary components for transcription.

A DNA template containing the desired genetic sequence is prepared. The templates are derived from DNA libraries designed in the first step of the mRNA display.

Template DNA Preparation

As the RNA polymerase moves along the DNA template, it incorporates the appropriate nucleotides to create a complementary RNA molecule. such as flow cytometry and ELISA.

RNA Synthesis

Transcription Reaction

In a reaction mixture, the DNA template combines with RNA polymerase enzymes, nucleotide units (A, U, G, C), and essential components.

Termination

Transcription persists until the RNA polymerase encounters a stop signal or reaches the terminus of the DNA template.

CD BioSciences' mRNA display technology provides robust support for your research endeavors. For more information or to inquire about service details, feel free to contact us at any time. They will be happy to assist you and tailor a solution that fits your project requirements.

Reference

Perenkov, A.D., et al. In vitro transcribed RNA-based platform vaccines: past, present, and future. Vaccines. 2023, 11(10): 1600.

For Research Use Only. Not For Clinical Use.

In Vitro Transcription

Principles of In Vitro Transcription

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