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Evaluation of Engineering Potential in Undomesticated Microbes With VECTOR.
Riley Williamson1, Nicholas Dusek2, Eglantina Lopez-Echartea1
1Department of Microbiological Sciences, North Dakota State University, Fargo, North Dakota, USA.
Microbial Biotechnology
|August 11, 2025
Summary
We developed VECTOR, a high-throughput pipeline to engineer diverse microbes beyond model organisms. This method enables scalable genetic modification of previously uncharacterized bacteria from complex communities.
Area of Science:
- Microbiology
- Synthetic Biology
- Genomics
Background:
- Genetic engineering is limited to well-characterized microbes like E. coli and B. subtilis.
- High-throughput culturomics yields large microbial collections, necessitating new engineering tools.
Purpose of the Study:
- To develop a scalable, high-throughput pipeline for assessing microbial engineerability.
- To enable genetic engineering of diverse, undomesticated microorganisms.
Main Methods:
- Developed VECTOR (Versatile Engineering and Characterisation of Transferable Origins and Resistance) pipeline.
- Utilized a library of tagged plasmids (BEVA architecture) with various origins of replication and antibiotic resistance genes.
- Delivered plasmids en masse to microbial communities and monitored transfer using fluorescence and sequencing.
Main Results:
- Successfully monitored genetic cargo transfer in real-time using OD600 and fluorescence.
- Identified optimal vector architectures for specific bacterial strains via plasmid barcode sequencing.
- Enabled isolation of engineered microbes from complex communities.
Conclusions:
- VECTOR provides a scalable solution for engineering diverse microbial communities.
- This platform expands the scope of genetic engineering beyond traditional model organisms.
- Facilitates the study and application of undomesticated microbes.
