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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Updated: Oct 15, 2025

Generation of Marked and Markerless Mutants in Model Cyanobacterial Species
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A CRISPR-Based Method for Constructing Conditional Mutations of Essential Genes in Cyanobacteria.

Ju-Yuan Zhang1, Tian-Cai Niu2, Gui-Ming Lin2

  • 1State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei Province, China. zhangjuyuan@ihb.ac.cn.

Methods in Molecular Biology (Clifton, N.J.)
|October 28, 2021
PubMed
Summary

Cyanobacteria research is advanced by a new CRISPR-based method. This technique allows conditional mutants of essential genes, overcoming previous genetic manipulation challenges in these photosynthetic bacteria.

Keywords:
CRISPRConditional mutantCyanobacteriaEssential gene

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Cyanobacteria are vital microorganisms for photosynthesis and nitrogen fixation.
  • They hold potential as cell factories for sustainable biosynthesis.
  • Genetic manipulation in cyanobacteria, especially for essential genes, is challenging due to limited tools.

Purpose of the Study:

  • To develop a novel method for creating conditional mutants of essential genes in cyanobacteria.
  • To overcome limitations in genetic manipulation of cyanobacteria.

Main Methods:

  • A CRISPR-based system was engineered.
  • Ribosome binding sites were modified using a theophylline-responsive riboswitch.
  • This facilitates conditional gene expression control.

Main Results:

  • A new method for constructing conditional mutants of cyanobacterial essential genes was successfully developed.
  • The method utilizes a theophylline-responsive riboswitch to control gene expression.
  • This approach addresses a significant bottleneck in cyanobacterial genetics.

Conclusions:

  • The developed CRISPR-based method enhances genetic manipulation in cyanobacteria.
  • This technique provides a powerful tool for studying essential genes and cellular functions.
  • The principle is broadly applicable to other bacterial species for conditional mutant construction.