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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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Targeted Nucleotide Editing Technologies for Microbial Metabolic Engineering.

Takayuki Arazoe1, Akihiko Kondo2,3, Keiji Nishida2

  • 1Department of Applied Biological Science, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan.

Biotechnology Journal
|June 5, 2018
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Summary
This summary is machine-generated.

CRISPR-Cas genome editing enables precise DNA modifications. New base editing technologies offer targeted nucleotide changes without DNA breaks, advancing microbial metabolic engineering.

Keywords:
CRISPR/Cas9base editinggenome editingmetabolic engineeringmicrobial engineering

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Area of Science:

  • Molecular Biology
  • Microbial Biotechnology
  • Genome Engineering

Background:

  • CRISPR-Cas systems provide programmable RNA-guided nucleases for genome editing.
  • DNA double-strand breaks (DSBs) induced by CRISPR-Cas are toxic to many microorganisms.
  • CRISPR-Cas is often used for negative selection in prokaryotes with other strategies.

Purpose of the Study:

  • To review and compare CRISPR-based genome editing technologies.
  • To discuss novel base editing platforms that avoid DSBs.
  • To explore applications in microbial metabolic engineering.

Main Methods:

  • Review of CRISPR-based genome editing techniques.
  • Comparison of current base editing platforms and their capabilities.
  • Analysis of nuclease-deficient CRISPR-Cas systems recruiting deaminases.

Main Results:

  • CRISPR-Cas systems facilitate gene disruptions, knockins, and translocations via DSB repair.
  • Novel base editing technologies enable targeted nucleotide conversions (C:G to T:A, A:T to G:C) without DSBs or donor DNA.
  • These advancements offer alternatives to DSB-inducing methods in prokaryotes.

Conclusions:

  • Base editing represents a significant advancement over traditional CRISPR-Cas editing for certain applications.
  • These technologies can overcome cellular regulation barriers in prokaryotes.
  • CRISPR-based genome editing and base editing are powerful tools for microbial metabolic engineering.