A universal molecular control for DNA, mRNA and protein expression

Area of Science:

• Molecular Biology
• Synthetic Biology
• Genomics
• Transcriptomics
• Proteomics

Background:

• Gene expression involves transcription to mRNA and translation to protein.
• Next-generation sequencing and mass spectrometry profile DNA, RNA, and protein abundance.
• A lack of universal reference standards hinders integrated multi-omics measurements.

Purpose of the Study:

• To engineer a universal molecular standard for multi-omics analysis.
• To create a control compatible with genomic, transcriptomic, and proteomic methods.
• To provide integrated gene expression measurement across molecular levels.

Main Methods:

• Utilized synthetic biology principles to engineer the pREF control.
• pREF encodes 21 synthetic genes for in vitro transcription and translation.
• Generated spike-in mRNA and matched protein controls from synthetic genes.

Main Results:

• pREF functions as a universal standard for next-generation sequencing and mass spectrometry.
• Synthetic genes provide qualitative controls for sensitivity and quantitative accuracy.
• Demonstrated pREF utility in metagenome DNA sequencing, RNA sequencing, and protein quantification.

Conclusions:

• pREF enables integrated measurement of gene expression across genomic, transcriptomic, and proteomic levels.
• The synthetic standard is independently propagable and sustainably prepared by laboratories.
• pREF addresses the need for compatible reference standards in multi-omics research.