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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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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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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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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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OptoCRISPRi-HD: Engineering a Bacterial Green-Light-Activated CRISPRi System with a High Dynamic Range.

Ke-Ning Chen1, Bin-Guang Ma1

  • 1Hubei Key Laboratory of Agricultural Bioinformatics, College of Informatics, Huazhong Agricultural University, Wuhan 430070, China.

ACS Synthetic Biology
|May 22, 2023
PubMed
Summary
This summary is machine-generated.

We developed a green-light-activated CRISPR interference (CRISPRi) system for precise gene regulation in E. coli. This optoCRISPRi-HD system offers a 40-fold dynamic range, enabling efficient control over essential and nonessential genes.

Keywords:
CRISPRibioprintingdynamic rangegene regulationoptogeneticssynthetic biology

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Microbiology

Background:

  • Gene expression modulation is vital for understanding gene function and engineering cellular behaviors.
  • Optogenetics and CRISPR interference (CRISPRi) offer precise control over gene expression.
  • Previous optoCRISPRi systems had limited dynamic range (≤10-fold) due to leakage, restricting applications.

Purpose of the Study:

  • To develop a high-dynamic-range, light-activated CRISPRi system for precise gene regulation in Escherichia coli.
  • To overcome the limitations of existing optoCRISPRi tools, particularly leakage activity.
  • To create a versatile tool for studying gene networks and cellular processes.

Main Methods:

  • Engineered a green-light-activated CRISPRi system in E. coli.
  • Achieved high dynamic range (40-fold) by minimizing leakage activity.
  • Demonstrated target gene repression and inhibition of DNA replication initiation.

Main Results:

  • The optoCRISPRi-HD system achieved a 40-fold dynamic range for gene repression.
  • Successfully repressed essential genes, nonessential genes, and inhibited DNA replication initiation.
  • Showcased spatial and temporal control over gene expression in E. coli.

Conclusions:

  • The developed optoCRISPRi-HD system provides a powerful tool for precise, light-inducible gene regulation in E. coli.
  • This system's high dynamic range and flexibility facilitate research in complex gene networks, metabolic engineering, and bioprinting.
  • Enables advanced applications requiring fine-tuned control over cellular functions.