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Engineering cell sensing and responses using a GPCR-coupled CRISPR-Cas system.

Nathan H Kipniss1, P C Dave P Dingal1, Timothy R Abbott1

  • 1Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.

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|December 22, 2017
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Summary
This summary is machine-generated.

Researchers developed a novel molecular device linking G-protein-coupled receptors (GPCRs) to CRISPR-dCas9 for precise genome regulation. This system enables cellular functions to be engineered by sensing diverse extracellular signals.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • G-protein-coupled receptors (GPCRs) are a vast family of eukaryotic membrane receptors sensing numerous extracellular cues.
  • Genome regulation is crucial for controlling cellular functions and responses.

Purpose of the Study:

  • To create a molecular device that couples CRISPR-dCas9 genome regulation to GPCRs.
  • To engineer a system for rewiring diverse ligand sensing to targeted genome regulation.

Main Methods:

  • Developed and compared two fusion architectures (Tango and ChaCha) for linking CRISPR to GPCRs.
  • Utilized mathematical modeling to analyze the CRISPR ChaCha design.
  • Tested the system's dose-dependency, reversibility, and ability to activate multiple genes.

Main Results:

  • The CRISPR ChaCha design allows for multiple dCas9 releases over a GPCR's lifetime.
  • The system is dose-dependent and reversible.
  • Successfully demonstrated activation of multiple endogenous genes in response to various ligands (synthetic compounds, chemokines, hormones, etc.).

Conclusions:

  • Introduced a modular toolkit of CRISPR-coupled GPCRs for versatile genome regulation.
  • This platform enables engineering cellular functions by linking ligand sensing to targeted gene activation.