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Precise Interference of RNA-Protein Interaction by CRISPR-Cas13-Mediated Peptide Competition

Meng Li1, Dan Li1, Leiruo Lin1

  • 1Molecular Cancer Research Center, School of Medicine, Shenzhen Campus of Sun Yat-sen University, Sun Yat-sen University, Shenzhen 518107, China.

ACS Synthetic Biology
|September 14, 2023
PubMed

Insights

Researchers developed a new CRISPR-based tool to precisely disrupt specific RNA-protein interactions. This method allows for a deeper understanding of how these interactions regulate gene expression and cellular processes in health and disease.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biotechnology

Background:

  • RNA-protein interactions are crucial for cellular regulation but their specific roles are often unclear.
  • The protein TTP binds to AU-rich elements (AREs) on mRNA via its TZF domain, repressing gene expression.

Purpose of the Study:

  • To develop a novel tool for precise interference of endogenous RNA-protein interactions.
  • To investigate the specific functions of RNA-binding protein (RBP) interactions in cellular processes.

Main Methods:

  • Engineered the CRISPR-Cas13b system to target specific RNA molecules.
  • Utilized a guide RNA (gRNA) to direct the TZF domain of TTP to compete with endogenous TTP binding sites on target mRNAs.
  • Interfered with TTP interactions with PIM1 and IL-2 3' UTRs.

Main Results:

  • Demonstrated specific interference with TTP binding to the AREs of PIM1 and IL-2 mRNAs.
  • Showed that precise interference with the TTP-PIM1 interaction had a distinct effect on cell proliferation compared to broad transcriptome interference.

Conclusions:

  • Established a novel CRISPR-based tool for precise RNA-protein interaction interference.
  • This tool enables a deeper understanding of the specific roles of individual RNA-RBP interactions in cellular regulation and disease.
Keywords:
3′ UTRAU-rich elementCRISPRCas13RNARNA binding protein

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