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Updated: Aug 20, 2025

Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR

Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR

Published on: November 23, 2012

Markerless Genome Editing in Competent Streptococci.

Roger Junges¹, Rabia Khan¹, Yanina Tovpeko²

¹Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.

Methods in Molecular Biology (Clifton, N.J.)

|November 23, 2022

Summary

This study introduces a markerless genome editing protocol for Streptococcus bacteria using natural transformation. This method enhances gene editing efficiency and precision without introducing unwanted genetic markers.

Keywords:

CSP Competence Markerless mutagenesis Natural transformation Pheromone Streptococcus Streptococcus mutans Streptococcus pneumoniae XIP

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Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning

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Published on: October 23, 2017

Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Classical mutagenesis often uses selective markers that can negatively impact gene expression and accumulate as unwanted genes.
Natural genetic transformation is a key mechanism for genetic exchange in bacteria like Streptococcus.

Purpose of the Study:

To develop a markerless genome editing protocol for Streptococcus mutans and Streptococcus pneumoniae.
To improve the efficiency and precision of gene editing via natural transformation.

Main Methods:

Utilizing natural transformation in Streptococcus species.
Inducing competence using specific peptides: sigX-inducing peptide pheromone (XIP) for S. mutans and competence-stimulating peptide (CSP) for S. pneumoniae.
Employing donor amplicons with extensive flanking homology for precise allelic integration.

Main Results:

Achieved high yields of transformants in both S. mutans and S. pneumoniae.
Demonstrated efficient and precise integration of new alleles without selectable markers.
Established a markerless genome editing system for these bacterial species.

Conclusions:

The developed protocol offers an efficient and precise method for markerless genome editing in Streptococcus species.
This approach overcomes limitations associated with traditional selective markers in mutagenesis.
Facilitates precise genetic manipulation for research and potential therapeutic applications.