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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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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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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Emergent CRISPR-Cas-based technologies for engineering non-model bacteria.

Daniel C Volke1, Enrico Orsi1, Pablo I Nikel1

  • 1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kongens Lyngby, Denmark.

Current Opinion in Microbiology
|July 6, 2023
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Summary
This summary is machine-generated.

CRISPR-Cas technologies revolutionize bacterial genome editing, making more microbes easier to engineer. This review explores using these tools for cell factory design and biotechnological applications.

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Area of Science:

  • Microbiology
  • Biotechnology
  • Molecular Biology

Background:

  • CRISPR-Cas systems have transformed bacterial genome engineering.
  • This has increased the genetic tractability of numerous non-model bacterial species.
  • These advancements are crucial for developing novel biotechnological applications.

Purpose of the Study:

  • To review recent trends in engineering non-model microbes using CRISPR-Cas technologies.
  • To discuss the potential of these engineered microbes in cell factory design.
  • To examine the role of CRISPR-Cas toolkits in enabling new biotechnological processes.

Main Methods:

  • Review of recent scientific literature on CRISPR-Cas applications in non-model bacteria.
  • Analysis of genome modification and transcriptional regulation strategies.
  • Examination of case studies in biotechnological process development.

Main Results:

  • CRISPR-Cas technologies enable precise genome modifications and tunable transcriptional control in non-model bacteria.
  • These tools facilitate the engineering of microbes for novel metabolic pathways, such as one-carbon substrate assimilation.
  • The development of CRISPR-Cas toolkits has expanded the scope of synthetic biology and metabolic engineering.

Conclusions:

  • CRISPR-Cas based genome engineering is a powerful approach for domesticating non-model bacteria.
  • These advancements are key to unlocking the potential of microbial cell factories for diverse biotechnological applications.
  • The future of bacterial genome engineering lies in harnessing the expanding capabilities of CRISPR-Cas systems.