pHaloFM Large-Fragment Editing

Insert up to 8 kb

A large-fragment genome editing tool designed for Halomonas, addressing genetic manipulation bottlenecks in non-model strains.

Yu-Hang, Zhang · Yuan, Yuan · Bai, Chen · Ya-Nan, Yang · Chen, Jiang · Lin, Tong · Wang, Shu · Zhu, Chen · Chen, Liang

Metabolic Engineering 2026

Specifications

Maximum insertion length

{'zh': '8', 'en': '8'} kb

Maximum deletion length

{'zh': '50', 'en': '50'} kb

Number of base editing targets

{'zh': '9', 'en': '9'}

Advantages

Insert up to 8 kb in one step

pHaloFM leverages native homologous recombination and a counterselection marker, enabling sequential insertion of multiple large fragments without multiple rounds of screening.

Delete up to 50 kb

The same plasmid system achieves deletions of up to 50 kb, facilitating genome minimization or removal of large regulatory regions.

Multiplex base editing

pHaloBE, adapted for the host, can edit nine target sites simultaneously, greatly accelerating construction of multigenic traits.

PHA copolymers produced

The tools were applied to construct complete biosynthetic pathways for P34HB and PHBV in Halomonas, demonstrating practical utility.

Applications

• Synthetic biology：Provides large-fragment tools for non-model strains like Halomonas, expanding chassis engineering.
• Biomanufacturing：Accelerates pathway construction in industrial strains; used to produce PHA copolymers for bioplastics.
• Microbial engineering：Enables one-step morphology and PHA pathway engineering, streamlining microbial engineering workflows.