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Transformation

Transformation

Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...

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Updated: Apr 28, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

Multiplex genome editing by natural transformation.

Ankur B Dalia¹, EmilyKate McDonough¹, Andrew Camilli²

¹Department of Molecular Biology and Microbiology and.

Proceedings of the National Academy of Sciences of the United States of America

|June 4, 2014

Summary

This summary is machine-generated.

Multiplex Genome Editing by Natural Transformation (MuGENT) enables rapid, scarless bacterial genome editing at high frequencies. This method accelerates evolution and genetic studies in naturally competent microbes.

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Area of Science:

Microbiology
Synthetic Biology
Genomics

Background:

Bacterial genome editing is crucial for research and synthetic biology.
Existing methods can be limited in efficiency and multiplexing capabilities.

Purpose of the Study:

To introduce and validate Multiplex Genome Editing by Natural Transformation (MuGENT) for accelerated evolution.
To demonstrate MuGENT's effectiveness in complex genome engineering and genetic interaction studies.

Main Methods:

Utilizing natural cotransformation of unlinked genetic markers in naturally competent microorganisms.
Employing DNA substrates with randomized nucleotides for unbiased editing.
Applying MuGENT for combinatorial genome editing (deletions, promoter replacements) and translation initiation tuning.

Main Results:

Achieved scarless genome editing at frequencies up to ~50%.
Demonstrated unbiased natural cotransformation suitable for directed evolution.
Generated complex mutant pools in one week, enabling a 30-fold improvement in natural transformation in Vibrio cholerae.
Showcased MuGENT's applicability in Streptococcus pneumoniae.

Conclusions:

MuGENT is a broadly applicable platform for accelerated evolution in naturally competent species.
The method facilitates complex multiplex genome editing and genetic interaction analysis.
MuGENT significantly enhances the speed and efficiency of bacterial strain engineering.