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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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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RNA Editing02:23

RNA Editing

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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.
The recognition sites for Cre recombinase called LoxP...
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Homologous Recombination02:31

Homologous Recombination

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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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CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans
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Reprogramming Acetogenic Bacteria with CRISPR-Targeted Base Editing via Deamination.

Peng-Fei Xia1, Isabella Casini1, Sarah Schulz1

  • 1Environmental Biotechnology Group, Center for Applied Geosciences, University of Tübingen, 72074 Tübingen, Germany.

ACS Synthetic Biology
|July 2, 2020
PubMed
Summary

A new base editing tool enables precise genome editing in acetogenic bacteria, facilitating the engineering of these microbes for biotechnology applications like improved chemical production.

Keywords:
CRISPRClostridium ljungdahliiacetogenic bacteriabase editingsynthetic biology

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Area of Science:

  • Microbiology
  • Synthetic Biology
  • Biotechnology

Background:

  • Acetogenic bacteria are valuable for converting C1 gases into organic chemicals.
  • Synthetic biology tools are crucial for engineering and understanding acetogenic bacteria.

Purpose of the Study:

  • To develop a precise genome-editing tool for acetogenic bacteria.
  • To engineer *Clostridium ljungdahlii* for enhanced acetate production.

Main Methods:

  • Developed a CRISPR-targeted deamination base-editing tool (nuclease-deactivated Cas9 and activation-induced cytidine deaminase).
  • Validated the tool in *Clostridium ljungdahlii* for cytosine-to-thymine substitutions.
  • Disrupted genes involved in acetate and ethanol production to redirect carbon flux.

Main Results:

  • Successfully implemented base editing for one-nucleotide resolution genome modification in acetogenic bacteria.
  • Engineered *C. ljungdahlii* strains exhibited improved acetate production with stable genotypes.
  • Identified potential for off-target edits through *in-silico* analysis.

Conclusions:

  • The developed base-editing tool advances genome engineering in acetogenic bacteria.
  • This technology provides a foundation for broader applications of CRISPR-Cas systems in bacterial research.
  • Reprogramming carbon flux enhances targeted chemical production in engineered microbial chassis.