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Related Experiment Video

Updated: Jul 1, 2026

High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii
10:51

High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii

Published on: December 5, 2016

An optimized, chemically regulated gene expression system for Chlamydomonas.

Paola Ferrante1, Claudia Catalanotti, Giulia Bonente

  • 1Italian National Agency for New Technologies, Energy and the Environment (ENEA), Casaccia Research Center, Rome, Italy.

Plos One
|September 13, 2008
PubMed
Summary

Researchers improved the Chlamydomonas CYC6 promoter for better gene expression control. Enhancements include increased induction strength and reversibility, enabling precise, repeatable gene expression "waves" for biotechnology.

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Published on: August 8, 2019

Area of Science:

  • * Molecular biology
  • * Algal biotechnology
  • * Gene expression regulation

Background:

  • * *Chlamydomonas reinhardtii* is a key model organism in algal and cell biology.
  • * It is utilized in biotechnological applications like molecular farming and biological hydrogen production.
  • * The metal-responsive CYC6 promoter in *Chlamydomonas* is typically induced by nickel and repressed by copper, but faces limitations in induction strength, reversibility, and potential toxicity.

Purpose of the Study:

  • * To enhance the strength and reversibility of the *Chlamydomonas* CYC6 promoter.
  • * To develop a more robust chemically regulated gene expression system for biotechnological applications.
  • * To overcome limitations of nickel induction, including weak induction, poor reversibility, and toxicity.

Main Methods:

  • * Utilized a codon-optimized *Renilla* luciferase reporter gene to assess promoter performance.
  • * Investigated the impact of the RBCS2 gene's first intron and a modified TAP medium on CYC6 promoter induction.
  • * Explored the use of copper chelators, specifically TETA (triethylenetetramine), as alternative inducers.

Main Results:

  • * Incorporating the RBCS2 intron or using modified TAP medium increased CYC6 promoter induction strength up to 20-fold.
  • * TETA demonstrated efficient induction at low concentrations and was more effective than nickel at similar levels.
  • * TETA-induced gene expression was fully reversible by copper, creating transient
  • Conclusions: [
  • * A finely tunable, chemically regulated gene expression system was developed for *Chlamydomonas*.
  • * The system allows for temporally controlled
  • * The optimized CYC6 promoter, combined with modified media, offers a reliable and sustainable method for controlled foreign gene expression in *Chlamydomonas*.

Conclusions:

  • * A finely tunable, chemically regulated gene expression system was developed for *Chlamydomonas*.
  • * The system allows for temporally controlled
  • * The optimized CYC6 promoter, combined with modified media, offers a reliable and sustainable method for controlled foreign gene expression in *Chlamydomonas*.