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| Autores principales: | , , , , , , |
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| Formato: | Artículo científico |
| Lenguaje: | en |
| Publicado: |
ACS synthetic biology
2026
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| Materias: | |
| Acceso en línea: | https://pubmed.ncbi.nlm.nih.gov/41979903/ |
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| _version_ | 1868266061970800640 |
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| author | Stuckless, Emily E Gai, Lian S Slattery, Samuel S Dempsey, Kira H Browne, Tyler S Gloor, Gregory B Edgell, David R |
| author_facet | Stuckless, Emily E Gai, Lian S Slattery, Samuel S Dempsey, Kira H Browne, Tyler S Gloor, Gregory B Edgell, David R Stuckless, Emily E Gai, Lian S Slattery, Samuel S Dempsey, Kira H Browne, Tyler S Gloor, Gregory B Edgell, David R |
| collection | PubMed - marine biology |
| 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. |
| format | Artículo científico |
| id | pubmed_41979903 |
| institution | PubMed |
| language | en |
| publishDate | 2026 |
| publisher | ACS synthetic biology |
| record_format | pubmed |
| spellingShingle | 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 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. |
| title | PHYCUT: Scalable Multiplex CRISPR/Cas9 Editing for Genome Engineering in the Diatom . |
| topic | Diatoms CRISPR-Cas Systems Gene Editing Glycosylation Plasmids Fucose |
| url | https://pubmed.ncbi.nlm.nih.gov/41979903/ |