skip to primary navigationskip to content

Reliable IP-free system for inter-chassis transfer of the high molecular weight DNA

This project aims to develop an IP-free system for transfer of the high molecular weight DNA from E. coli to M. polymorpha. This includes development of the plant-specific iBACs for the reliable transfer and integration of the high molecular weight DNA into the M. polymorpha genome. Alternative methods for interchassis DNA transfer, such as integrative and conjugative elements (ICEs) will be also explored.

The Idea

The ability to manipulate and transfer the high molecular weight DNA molecules between different chassis constitutes one of the main bottlenecks of the rational genome engineering. Bacterium Escherichia coli and plant Marchantia polymorpha are both well-characterized model organisms that are frequently used as chassis for biotechnology and synthetic biology applications. Although the vast majority of the good DNA assembly and editing tools are in E. coli, M. polymorpha is better host for certain applications. Novel tools combining the benefits of M. polymorpha chassis and the reliable DNA recombineering approaches developed for E. coli are critical for developing a robust synthetic biology toolkit. Bacterial artificial chromosomes (BACs) based on the E. coli fertility factor (F-factor) are frequently used for engineering high molecular weight DNA fragments in E. coli. We have recently engineered integrative bacterial artificial chromosomes (iBACs) that can accept virtually any high molecular weight DNA fragment for integration into B. subtilis chromosome and allow rapid selection of transformants by B. subtilis-specific antibiotic resistance and the yellow fluorescent protein (mVenus) expression. This project aims to develop an IP-free system for transfer of the high molecular weight DNA from E. coli to M. polymorpha. This includes development of the plant-specific iBACs for the reliable transfer and integration of the high molecular weight DNA into the M. polymorpha genome. Alternative methods for interchassis DNA transfer, such as integrative and conjugative elements (ICEs) will be also explored.

 

The Team

Dr Mario Juhas
Generic Avatar
Dave Willey
Christian Boehm

 

 

 

 

 

 

 

Dr. Mario Juhas

Research Associate, Department of Pathology

Dr. Dave Willey

Christian R. Boehm

PhD Candidate in Plant Synthetic Biology, Department of Plant Sciences

The project will benefit from the collaboration with the Jim Haseloff's lab at the Department of Plant Sciences of the University of Cambridge with expertise in the M. polymorpha genome modification and analysis

Collaborator from the Faculty of Law might join the project later (for IP related issues)

 

Project Outputs

Filed under:

About

The Synthetic Biology Strategic Research Initiative provides a hub for anyone interested in Synthetic Biology at the University of Cambridge, including researchers, commercial partners and external collaborators. 

Subscribe to our mailing list