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The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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Chromosomal Looping-Based Expression Activation System in Yeast.

Leilei Qiu1, Xiaoying Wang1, Gui Zhuang1

  • 1State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, 361102, China.

Small Methods
|December 20, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed CLEASY, a novel tool for engineering DNA looping in yeast. This system enables controllable gene expression by manipulating chromosomal architecture, offering a simplified eukaryotic model for studying gene regulation.

Keywords:
DNA loopinggene expressionphotoreceptorphytohormoneyeast

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Area of Science:

  • Molecular Biology
  • Genomics
  • Synthetic Biology

Background:

  • 3D genome organization is a key regulator of gene expression.
  • Existing tools for engineering chromosomal looping in simple eukaryotes are limited.
  • Controllable manipulation of DNA interactions is crucial for understanding gene regulation.

Purpose of the Study:

  • To develop a novel tool for engineering chromosomal looping and gene expression in yeast.
  • To create a simplified eukaryotic system for studying DNA looping mechanisms.
  • To establish a platform for investigating molecules that influence looping and gene activation.

Main Methods:

  • Development of the chromosomal looping-based expression activation system in yeast (CLEASY).
  • Utilizing conditionally interacting proteins, distal regulatory elements, and a reporter gene on a modified yeast chromosome.
  • Employing exogenous chemical or light induction to trigger protein interactions and gene activation.
  • Ensuring target specificity and variability using bivalent Cas9 complexes and guide RNAs.

Main Results:

  • Successful development of CLEASY, a controllable system for inducing DNA looping.
  • Demonstrated activation of a reporter gene through proximity of distal regulatory elements.
  • Confirmed compatibility of CLEASY with Cas9 systems for targeted gene regulation.

Conclusions:

  • CLEASY provides a simplified eukaryotic model for engineering DNA looping machinery.
  • The system offers a controllable and specific method for manipulating gene expression.
  • CLEASY serves as a valuable platform for investigating looping mechanisms and identifying effective molecules.