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CRISPR and crRNAs02:53

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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 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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Microbial growth control refers to various methods employed to inhibit, reduce, or eliminate microorganisms to ensure safety and hygiene across different settings. These methods are categorized based on the target environment and the level of microbial control required.Biocides are versatile agents designed to control microorganisms by either inhibiting their growth or outright killing them. These agents work through various physical, chemical, mechanical, or biological mechanisms. The...
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Related Experiment Video

Updated: Feb 28, 2026

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

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Controlling microbial PHB synthesis via CRISPRi.

Dan Li1, Li Lv1, Jin-Chun Chen1

  • 1Center for Synthetic and Systems Biology, School of Life Science, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, 100084, China.

Applied Microbiology and Biotechnology
|June 17, 2017
PubMed
Summary
This summary is machine-generated.

CRISPR interference (CRISPRi) precisely controlled polyhydroxyalkanoates (PHA) synthesis in E. coli by regulating PHA synthase activity. This method enabled tunable PHA content and molecular weight for advanced material applications.

Keywords:
CRISPRiCas9Molecular weightPHA synthasePHBPhaCPolyhydroxyalkanoatesSynthetic biology

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Polymer Science

Background:

  • Microbial polyhydroxyalkanoates (PHA) are biopolyesters with properties comparable to petroleum plastics.
  • Polyhydroxybutyrate (PHB) is the most prevalent type of PHA.
  • CRISPR interference (CRISPRi) offers a novel method for gene expression control in various organisms.

Purpose of the Study:

  • To investigate the use of CRISPRi to regulate PHA synthase activity and influence PHB synthesis in recombinant Escherichia coli.
  • To determine the impact of CRISPRi-mediated gene regulation on PHB content, molecular weight, and polydispersity.

Main Methods:

  • Recombinant E. coli strains containing the phaCAB operon were engineered.
  • Single-guide RNAs (sgRNAs) were designed to target the PhaC synthase gene for CRISPRi.
  • CRISPRi was employed to modulate phaC transcription and PhaC activity.

Main Results:

  • CRISPRi effectively controlled phaC transcription and PhaC activity based on sgRNA binding location and number.
  • PHB content showed a direct correlation with PhaC activity, while molecular weight and polydispersity exhibited inverse relationships.
  • PHB content was tunable from 1.47-75.21% cell dry weight, with molecular weights ranging from 2 to 6 million Daltons.

Conclusions:

  • CRISPRi is a powerful tool for precisely regulating PHA biosynthesis pathways.
  • This study demonstrates the ability to control PHA properties, including molecular weight and content, opening avenues for high-strength bioplastic applications.