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bioRxiv · 10.1101/2025.07.11.664204

Combinatorial pathway engineering using multiplex Serine recombinase-Assisted Genome Engineering (mSAGE)

Huenemann, J. D.·Carroll, A. L.·Elmore, J. R.·Dexter, G.·Mand, T.·Klingeman, D. M.·Alexander, W.·Guss, A. M.
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Abstract

Low efficiency of DNA integration into the chromosome limits multiplexed bacterial genetic engineering. We address this by establishing multiplexed Serine recombinase-Assisted Genome Engineering (mSAGE), enabling simultaneous, site-specific insertion of three DNAs into the chromosome of Pseudomonas putida KT2440. We construct a combinatorial library of isophthalate catabolism and transport genes through a single, pooled transformation of three gene libraries, rapidly identifying the best combination of genes for biodegradation of this plastic comonomer.

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Huenemann, J. D., Carroll, A. L., Elmore, J. R., Dexter, G., Mand, T., Klingeman, D. M., Alexander, W., Guss, A. M.. 2025-07-11. Combinatorial pathway engineering using multiplex Serine recombinase-Assisted Genome Engineering (mSAGE). https://doi.org/10.1101/2025.07.11.664204

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