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RNA Editing02:23

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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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Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
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DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their  survival. Therefore, the copying errors are checked and repaired at three levels.
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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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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
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Updated: Aug 15, 2025

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
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Cytosine Base Editing in Bacteria.

Ye Liu1,2, Yang Liu1,2, Ping Zheng1,2

  • 1Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.

Methods in Molecular Biology (Clifton, N.J.)
|January 2, 2023
PubMed
Summary
This summary is machine-generated.

Cytosine base editing offers a precise way to alter bacterial DNA without breaking it. This chapter details a reliable protocol for base editing in Corynebacterium glutamicum and Bacillus subtilis.

Keywords:
Bacillus subtilisBase editingCRISPRCorynebacterium glutamicumCytidine deaminaseGenome editing

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Area of Science:

  • Molecular Biology
  • Genetics
  • Microbial Biotechnology

Background:

  • Base editing is an advanced genome editing technique.
  • It allows precise DNA base modifications without DNA cleavage or donor templates.
  • It is widely applied in engineering eukaryotic and prokaryotic microorganisms.

Purpose of the Study:

  • To describe a routine protocol for cytosine base editing.
  • To demonstrate its application in two model bacteria: Corynebacterium glutamicum and Bacillus subtilis.

Main Methods:

  • A routine protocol for cytosine base editing was developed.
  • The protocol was applied to Corynebacterium glutamicum and Bacillus subtilis.

Main Results:

  • The protocol provides a reliable method for base editing in the selected bacterial models.
  • The method enables targeted DNA base mutations.

Conclusions:

  • The described protocol is effective for cytosine base editing in Corynebacterium glutamicum and Bacillus subtilis.
  • The protocol can be adapted for base editing in other bacterial species with necessary modifications.