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PERSIST platform provides programmable RNA regulation using CRISPR endoRNases.

Breanna DiAndreth1,2, Noreen Wauford1,2, Eileen Hu1,2,3

  • 1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.

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|May 13, 2022
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Summary

Researchers developed a new RNA-regulation platform called PERSIST to overcome gene silencing issues in gene therapies. This system offers robust control over transgene expression, enhancing reliability for advanced therapeutic applications.

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Gene Therapy

Background:

  • Transcription factor-based gene regulation is crucial for gene therapies but faces limitations like epigenetic silencing.
  • Existing methods struggle with expression longevity and reliability due to silencing, necessitating alternative regulatory strategies.

Purpose of the Study:

  • To develop a novel RNA-regulation platform to achieve robust and long-lasting transgene expression.
  • To create a system that overcomes the limitations of transcription factor-based regulation, specifically epigenetic silencing.

Main Methods:

  • Development of the PERSIST platform, utilizing nine CRISPR-specific endoRNases as RNA-level activators and repressors.
  • Incorporation of modular OFF- and ON-switch regulatory motifs within the PERSIST system.
  • Testing of PERSIST-regulated transgenes for OFF/ON responses, silencing resistance, and circuit construction capabilities.

Main Results:

  • PERSIST-regulated transgenes demonstrated strong OFF and ON responses.
  • The platform successfully resisted gene silencing for a minimum of two months.
  • PERSIST enabled the construction of complex gene circuits, including cascades and logic functions.

Conclusions:

  • The PERSIST platform offers an orthogonal, modular, and composable solution for gene regulation.
  • This RNA-level regulation system provides robust and predictable gene circuit construction.
  • PERSIST holds significant promise for diverse applications in gene and cell therapies.