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Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays

Published on: November 12, 2012

A CRISPR-based chromosomal-separation technique for Escherichia coli.

Junchang Su^1,2, Pengju Wang^2,3,4, Ju Li⁵

¹School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, China.

Microbial Cell Factories

|November 11, 2022

Summary

Scientists engineered multi-chromosomal Escherichia coli (E. coli) strains using a novel CRISPR-based method. This genome engineering technique creates new E. coli strains with split chromosomes, advancing synthetic biology applications.

Keywords:

CRISPR-Cas9 Chromosomal-separation Genome engineering Synthetic biology

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A Method to Study de novo Formation of Chromatin Domains

A Method to Study de novo Formation of Chromatin Domains

Published on: August 23, 2019

Area of Science:

Microbiology
Genomics
Synthetic Biology

Background:

Genome engineering enables significant modification of natural life systems.
Previous methods for creating large DNA structures in E. coli were often inconvenient or required engineered strains.

Purpose of the Study:

To develop a novel, simple, and efficient method for creating multi-chromosomal strains of wild-type Escherichia coli.
To demonstrate the feasibility of splitting the native E. coli chromosome without using heterologous genetic parts.

Main Methods:

Utilized a CRISPR-based genome engineering technique to split the native chromosome of wild-type E. coli.
Verified the new chromosomal arrangements using PCR amplification of joint regions and Nanopore/Illumina sequencing.
Generated two distinct E. coli strains with different chromosomal configurations: E. coli^0.10/4.54 and E. coli^2.28/2.36.

Main Results:

Successfully created novel E. coli strains with two chromosomes (0.10 Mb/4.54 Mb and 2.28 Mb/2.36 Mb).
Confirmed chromosomal splitting via sequencing and PCR analysis.
Observed that E. coli^0.10/4.54 was stable, while E. coli^2.28/2.36 underwent recombination. Both engineered strains exhibited slightly slower growth and elongated cell shapes compared to wild-type E. coli.

Conclusions:

A simple CRISPR-based genome engineering technique was successfully developed for constructing multi-chromosomal E. coli strains.
The developed technique does not require heterologous genetic parts.
This method holds potential for application in other prokaryotes for synthetic biology research and development.