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Related Experiment Video

Updated: Oct 18, 2025

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
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Chimeric RNA-binding protein-based killing switch targeting hepatocellular carcinoma cells.

Jiong Yang1,2, Shigang Ding1,2

  • 1Department of Gastroenterology, Peking University Third Hospital, Beijing 100191, China.

Molecular Therapy. Nucleic Acids
|September 30, 2021
PubMed
Summary

Synthetic biology engineered RNA-binding proteins (RBPs) to create cancer-specific killing circuits. This new system effectively targets and eliminates hepatocellular carcinoma (HCC) cells in mixed cell populations.

Keywords:
HCCRNA-binding proteinmodRNA deliveryproteasesynthetic circuit

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Area of Science:

  • Synthetic biology
  • Molecular engineering
  • Cancer therapeutics

Background:

  • Synthetic circuits offer targeted cancer cell elimination.
  • Proteolysis-responding capsid-cNOT7s show promise but underperform with modified mRNA (modRNA) delivery.
  • Development of alternative modules for modRNA delivery is crucial for improving synthetic circuit efficacy.

Purpose of the Study:

  • To engineer novel RNA-binding proteins (RBPs) for enhanced modRNA-based synthetic circuits.
  • To develop and test chimeric RBPs compatible with proteolysis-based sensing units.
  • To construct a hepatocellular carcinoma (HCC) cell-specific killing circuit using the engineered RBP system.

Main Methods:

  • Engineering RBPs by fusing them with degron and cleavage sites.
  • Testing the compatibility of chimeric RBPs with proteolysis-based sensing units.
  • Implementing eight two-input and four three-input logic gates.
  • Constructing an HCC cell-specific killing circuit with proteolysis-based sensing units, a two-input OR gate, and an apoptosis-inducing actuator.

Main Results:

  • Successfully engineered chimeric RBPs compatible with proteolysis-based sensing units.
  • Implemented various logic gates (2-input and 3-input) using the chimeric RBP system.
  • Developed a functional HCC cell-specific killing circuit.
  • Demonstrated the circuit's ability to distinguish HCC cells and induce apoptosis in a mixed cell population.

Conclusions:

  • Engineered chimeric RBPs provide a viable alternative for modRNA-based synthetic circuits.
  • The developed RBP-based system enables the construction of sophisticated logic gates for biological computation.
  • The HCC cell-specific killing circuit effectively targets and eliminates cancer cells, showing potential for cancer therapy.