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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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A yeast-based rapid prototype platform for gene control elements in mammalian cells.

Kathy Y Wei1, Yvonne Y Chen, Christina D Smolke

  • 1Department of Bioengineering, Stanford University, 473 Via Ortega, MC 4201, Stanford, CA 94305, USA.

Biotechnology and Bioengineering
|November 28, 2012
PubMed
Summary

Researchers developed a plug-and-play ribozyme device platform for programming genetic circuits in mammalian cells. This system, transferable from yeast, enables rapid prototyping and high-throughput screening of gene controllers for diverse applications.

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Programming mammalian genetic circuits demands adaptable, user-specific gene-control systems.
  • Current systems often require extensive re-engineering for mammalian cell compatibility.

Purpose of the Study:

  • To demonstrate a novel ribozyme-based device platform for direct transfer and function in mammalian cells.
  • To establish a rapid prototyping platform (RPP) for developing mammalian gene controllers.

Main Methods:

  • Utilized ribozyme switches previously prototyped in yeast.
  • Tested device performance in human HEK 293, HeLa, and U2OS cell lines without sequence modification.
  • Assessed gene expression regulation, ligand responsiveness, and device function (ON/OFF switch).

Main Results:

  • Ribozyme devices functioned predictably and ligand-responsively in mammalian cells, mirroring yeast performance.
  • Activation ratios were comparable to existing RNA-based regulatory devices in mammalian cells.
  • Strong correlations (R(2) = 0.63-0.97) observed between yeast and mammalian cell performance.

Conclusions:

  • The ribozyme platform offers a "plug-and-play" solution for mammalian gene circuit design.
  • Yeast can serve as an effective chassis for rapid prototyping and screening of mammalian gene controllers.
  • This approach accelerates the development of gene controllers for cell-based therapeutics and cell-fate reprogramming.