A methylome-guided toolkit for genetic engineering of the fast-growing cyanobacterium UTEX 3222
- 1General Biotechnologies
Abstract
Cyanobacterium aponinum UTEX 3222 is a fast-growing marine cyanobacterium that reaches high cell densities, making it an attractive chassis strain for low-cost biomanufacturing. However, past efforts to genetically engineer UTEX 3222 have been unsuccessful. We hypothesized that the complex restriction modification systems in the UTEX 3222 genome constitute a major barrier to genetic transformation. Using a combined methylation avoidance and methylation mimicking approach, we overcame this barrier and successfully transformed UTEX 3222 by natural transformation, generating a large panel of fully segregated axenic strains. Transformation method was also established for a novel shuttle vector that replicates in UTEX 3222. A large-scale promoter screen identified promoters which exhibited strong expression at high cell density. These advances establish the foundation for developing UTEX 3222 as a scalable platform for photosynthetic biomanufacturing.
Related
- The genetic architecture of human DNA methylome variation
- A Yeast Logic Toolkit for Fast and Efficient Assembly of Genetic Circuits
- Engineering Persistent and Rewritable Genetic Memory in Bacteria through Multiscale Plasmid Competition
- Complement system in kidney transplantation: from innate immune surveillance to a therapeutic and genetic engineering target
- Baseline growth rate shapes pollutant impacts in aquatic environments: Evidence from antibiotic responses in the harmful cyanobacterium Microcystis
BibTeX
@misc{lin2026methylomeguided,
title = {{A methylome-guided toolkit for genetic engineering of the fast-growing cyanobacterium UTEX 3222}},
author = {D. Lin and A. Booth and A. Gill and M. M. Aung and D. Kim and U. S. Sagaram},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.15.751560},
url = {https://www.biorxiv.org/content/10.64898/2026.09.15.751560v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - A methylome-guided toolkit for genetic engineering of the fast-growing cyanobacterium UTEX 3222
AU - D. Lin
AU - A. Booth
AU - A. Gill
AU - M. M. Aung
AU - D. Kim
AU - U. S. Sagaram
PY - 2026
DA - 2026/09/15
PB - bioRxiv
DO - 10.64898/2026.09.15.751560
UR - https://www.biorxiv.org/content/10.64898/2026.09.15.751560v1
AB - Cyanobacterium aponinum UTEX 3222 is a fast-growing marine cyanobacterium that reaches high cell densities, making it an attractive chassis strain for low-cost biomanufacturing. However, past efforts to genetically engineer UTEX 3222 have been unsuccessful. We hypothesized that the complex restriction modification systems in the UTEX 3222 genome constitute a major barrier to genetic transformation. Using a combined methylation avoidance and methylation mimicking approach, we overcame this barrier and successfully transformed UTEX 3222 by natural transformation, generating a large panel of fully segregated axenic strains. Transformation method was also established for a novel shuttle vector that replicates in UTEX 3222. A large-scale promoter screen identified promoters which exhibited strong expression at high cell density. These advances establish the foundation for developing UTEX 3222 as a scalable platform for photosynthetic biomanufacturing.
ER -