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| Main Authors: | , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
ACS synthetic biology
2026
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/41979903/ |
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Table of Contents:
- PHYCUT: Scalable Multiplex CRISPR/Cas9 Editing for Genome Engineering in the Diatom . Stuckless, Emily E Gai, Lian S Slattery, Samuel S Dempsey, Kira H Browne, Tyler S Gloor, Gregory B Edgell, David R Diatoms CRISPR-Cas Systems Gene Editing Glycosylation Plasmids Fucose Diatoms are globally significant microalgae that contribute ∼20% of oxygen production and exhibit remarkable metabolic diversity. The marine diatom has emerged as a promising synthetic biology platform for the bioproduction of recombinant proteins, supported by a human-like -linked glycosylation pathway. However, its α(1,3)-linked core fucose is potentially immunogenic in humans and thus limits its biopharmaceutical applications. One hurdle to efficient genome engineering in is the lack of a robust system for simultaneous CRISPR/Cas9 editing at multiple sites. To overcome this limitation, we develop PHYCUT ( Cs4-as9 mltiplex ool), a versatile plasmid-based CRISPR/Cas9 system that uses the Csy4 endoribonuclease to process multiguide RNA arrays. To highlight PHYCUT applications, we demonstrate multiplex editing of all three genes responsible for α(1,3) fucosylation in , yielding strains with reduced fucosylation of secreted proteins. PHYCUT enables facile, multiplexed genome engineering in diatoms and provides a foundation for humanizing the glycosylation pathway to support next-generation algal biotechnology.