A common concern with mRNA display is the susceptibility of RNA to degradation. The technology developed by CD BioSciences effectively addresses this issue. CD BioSciences specializes in the development and application of mRNA display technology, providing valuable insights into the drug discovery process.

Fig 1. General scheme for peptide selection by mRNA display.

Reverse Transcription in mRNA Display

Reverse transcription is the process where the enzyme reverse transcriptase creates cDNA from an RNA template. In mRNA display, fusions are converted into cDNA either before or occasionally after the selection step. The cDNA from selected fusions is then amplified using PCR to regenerate the DNA template library for the next round of selection.

Fig 2. Advancement of mRNA display into cDNA display. (Newton, et al., 2019)

mRNA-dependent technology has been perceived as technically challenging due to the labile nature of RNA molecules. A common question about mRNA display involves the sensitivity of RNA to degradation. Converting mRNA to cDNA provides a new strategy to address this issue.

• The problem of the mRNA tag interfering with selection is mitigated by performing reverse transcription before selection. This creates an mRNA/cDNA hybrid, which helps prevent the formation of secondary structures.
• Reverse transcription to form an mRNA/cDNA duplex has been implemented to increase the stability of mRNA-peptide conjugates and reduce nonspecific interactions between mRNA and the target molecule.

Our Services

To ensure the continuation of mRNA display, CD BioSciences rapidly converts the unstable mRNA-protein fusion into a stable mRNA/cDNA-protein fusion before selection. The labile nature of the mRNA region has been improved by synthesizing its cDNA counterpart and generating an mRNA/cDNA-protein fusion molecule using a puromycin-linker DNA that harbors a primer for reverse transcription.

Although the literature reports that RNA nuclease degradation is not an issue due to the in vitro nature of the entire mRNA display procedure, we still recommend performing this step to minimize the possibility of degradation and eliminate experimental interference.

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CD BioSciences' mRNA display services are meticulously designed to meet your research needs with precision and excellence. Partner with us for unparalleled expertise and support in early drug discovery. contact us for more information.

Reference

1. Newton, M.S., et al. In vitro selection of peptides and proteins—advantages of mRNA display. ACS Synthetic Biology. 2019, 9(2): 181-190.

Related Services

• In Vitro Transcription
• In Vitro Translation
• Affinity Selection
• Analysis and Optimization
• DNA Library Preparation
• Peptide Cyclization
• Puromycin Ligation