Columbia Technology Ventures

High-throughput in vivo mRNA-based proteomics via next-generation sequencing

This technology is a platform for high-throughput, in vivo proteomic analysis that links expressed proteins to their corresponding RNA sequences using an RNA-binding domain and recognition elements, enabling precise identification via nucleic-acid-based sequencing.

Unmet Need: Scalable, accurate proteomic analysis in vivo

Current methods for proteomic analysis rely on mass spectrometry or antigen detection, which are limited by low throughput, high costs, specialized equipment, and reliance on in vitro conditions, which can obscure the true dynamics of the proteome in vivo. These approaches fail to provide scalable solutions for examining complex protein interactions and localization in vivo. As such, high-throughput, cost-effective, and accurate approaches for proteomic analysis in biological contexts remain limited.

The Technology: In vivo mRNA display for high-throughput proteomic analysis

This technology is an assay that utilizes mRNA display technology for high-throughput proteomic analysis by linking expressed proteins to their corresponding mRNA sequences. The system employs an mRNA display cassette containing an RNA-binding domain fused to the protein of interest, and a unique RNA stem-loop that enables high-affinity binding. This technique enables in vivo quantitative analysis of protein expression through nucleic acid sequencing, therefore preserving the native cellular environment. Additionally, barcoding mRNAs with unique molecular identifiers (UMIs) allows precise and scalable analysis of protein behaviors, offering significant advancement over traditional in vitro methods.

This technology has been validated in Saccharomyces cerevisiae, demonstrating the potential to enable high-throughput proteomic screening that can be adapted for other organisms.

Applications:

  • High-throughput analysis of protein expression and interactions
  • Quantitative protein affinity and binding dynamics measurements
  • Identification and validation of therapeutic targets
  • Variant protein characterization and subcellular localization
  • Characterization of protein binding domains
  • Protein engineering

Advantages:

  • Single-nucleotide resolution
  • Compatible across multiple eukaryotic cells
  • High throughput for large-scale analysis
  • Preserves native cellular context
  • Multiplexed measurements
  • Does not require specialized equipment
  • Cost-effective and scalable, reducing processing costs to one-tenth of spectrometry-based approaches

Lead Inventor:

Saeed Tavazoie, Ph.D.

Patent Information:

Patent Pending (US20230125614)

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