---
title: "BRIDGE: Broad-host interkingdom DNA delivery platform for genetic domestication of diverse yeasts"
id: "biorxiv-7-bridge-a-broad-host-platform-for-interkingdom-dna-delivery-enabling-the-genetic"
canonical_url: "https://medichelpline.com/clinical-feed/biorxiv-7-bridge-a-broad-host-platform-for-interkingdom-dna-delivery-enabling-the-genetic"
content_type: "clinical_feed_article"
specialty: "General"
source_name: "bioRxiv (Biomedical Preprints)"
source_url: "https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1?rss=1"
published_at: "2026-09-21T12:00:00.000Z"
evidence_level: "Verified Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# BRIDGE: Broad-host interkingdom DNA delivery platform for genetic domestication of diverse yeasts
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/biorxiv-7-bridge-a-broad-host-platform-for-interkingdom-dna-delivery-enabling-the-genetic
- **Specialty:** [General](https://medichelpline.com/clinical-feed/general.md)
- **Primary Source:** bioRxiv (Biomedical Preprints)
- **Source URL:** [Original Journal Publication](https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1?rss=1)
- **Published At:** 2026-09-21T12:00:00.000Z
- **Evidence Rating:** Verified Feed
## Executive GIST (TL;DR)
- BRIDGE is an integrated synthetic biology platform enabling broad-host **interkingdom DNA** delivery from bacteria to yeasts, addressing a key barrier to engineering non-conventional yeasts. - The system is built around a compact 6-kb **Pan/ARS-oriT broad-host BRIDGE vector** and uses a superconjugative helper plasmid, **pSC5**, to mediate transfer. - Using BRIDGE, interkingdom DNA transfer was extended from four genera accessible with pSC5 alone to ten phylogenetically diverse yeast genera, including **Saccharomyces**, Maudiozyma, Starmerella, Kazachstania, Yarrowia, Zygosaccharomyces, Lachancea, Kluyveromyces, Pichia and Komagataella. - To validate multigene transfer, the authors assembled a 16-kb visual reporter carrying a synthetic five-gene **violacein** biosynthetic pathway from Chromobacterium violaceum. - Construction of transcriptional units used the **YeastFab** modular cloning system; full pathway assembly was achieved by single-step in vivo homologous recombination in Saccharomyces cerevisiae, and plasmids were recovered via the **EASY-C** platform. - The 16-kb reporter was delivered successfully to all ten yeast genera, demonstrating transfer of complete multigene synthetic pathways across broad hosts. - Robust violacein production was observed across six Saccharomyces species tested, enabling rapid visual selection, while modest expression was seen in Kazachstania and Kluyveromyces; other genera received the pathway but showed no visible pigmentation. - The authors conclude that **regulatory compatibility**, rather than DNA transfer capacity, is the main determinant of heterologous pathway expression in distantly related yeasts. - BRIDGE establishes a Design-Build-Recover-Deliver workflow that combines modular in vitro transcriptional unit construction, single-step in vivo multigene assembly, rapid plasmid recovery and broad-host transfer, facilitating rapid genetic modification of previously intractable microorganisms. - The platform is presented as a tool to expand the synthetic biology toolbox for sustainable biomanufacturing and industrial biotechnology.
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Reem Swidah 1 The University of Manchester; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Reem%2BSwidah%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Swidah%20R&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AReem%2BSwidah%2B) * [ORCID record for Reem Swidah](http://orcid.org/0000-0003-2695-1931 "Open in new tab") * For correspondence: reem.swidah@manchester.ac.uk Ryan Cochrane 1 The University of Manchester; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Ryan%2BCochrane%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Cochrane%20R&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ARyan%2BCochrane%2B) * [ORCID record for Ryan Cochrane](http://orcid.org/0009-0004-5154-833X "Open in new tab") Fernando Valle 2 BP Biosciences Centre, San Diego, CA 92121, USA * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Fernando%2BValle%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Valle%20F&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AFernando%2BValle%2B) Daniela Delneri 1 The University of Manchester; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Daniela%2BDelneri%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Delneri%20D&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ADaniela%2BDelneri%2B) * [ORCID record for Daniela Delneri](http://orcid.org/0000-0001-8070-411X "Open in new tab") * [Abstract](https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_art/node:5799401/1) * [Info/History](https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1.article-info)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_info/node:5799401/1) * [Metrics](https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1.article-metrics)[](https://www.biorxiv.org/panels_ajax_tab/article_tab_metrics/node:5799401/1) * [Supplementary material](https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1.supplementary-material)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_data/node:5799401/1) * [ Preview PDF](https://www.biorxiv.org/content/10.64898/2026.09.17.752325v1.full.pdf+html)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_pdf/node:5799401/1) ![Loading](https://www.biorxiv.org/sites/all/modules/contrib/panels_ajax_tab/images/loading.gif) ## Abstract Efficient DNA delivery remains one of the greatest barriers to engineering non-conventional yeasts, limiting their genetic domestication and exploitation as next-generation microbial cell factories. Here, we present BRIDGE (Bacteria-to-yeast Rapid Interkingdom DNA Gene Exchange), an integrated synthetic biology platform for broad-host interkingdom DNA delivery. BRIDGE is centred on a compact 6-kb Pan/ARS-oriT broad-host BRIDGE vector, enabled by a superconjugative helper plasmid (pSC5). Using BRIDGE, we expanded interkingdom DNA transfer from representatives of four genera accessible using pSC5 alone to ten phylogenetically diverse yeast genera, including Saccharomyces, Maudiozyma, Starmerella, Kazachstania, Yarrowia, Zygosaccharomyces, Lachancea, Kluyveromyces, Pichia and Komagataella. To demonstrate the versatility of BRIDGE, we engineered a 16-kb visual reporter carrying a synthetic five-gene violacein biosynthetic pathway from Chromobacterium violaceum. Individual transcriptional units were first constructed using the YeastFab modular cloning system and subsequently assembled into the complete pathway by single-step in vivo homologous recombination in Saccharomyces cerevisiae, followed by rapid plasmid recovery using the EASY-C platform. The reporter was successfully delivered to all ten yeast genera, demonstrating BRIDGE-mediated transfer of complete multigene synthetic pathways. Robust violacein production was observed across all six Saccharomyces species tested, providing a rapid visual marker for transformant identification, while modest functional pathway expression was also detected in Kazachstania and Kluyveromyces. Successful pathway transfer, albeit without visible pigmentation, was achieved in the remaining genera, indicating that regulatory compatibility, rather than DNA transfer, is the principal determinant of heterologous pathway expression in distantly related yeasts. Collectively, BRIDGE establishes an integrated Design-Build-Recover-Deliver workflow that combines modular in vitro construction of transcriptional units, single-step in vivo assembly of multigene pathways, rapid plasmid recovery using the EASY-C platform and broad-host interkingdom DNA delivery. This versatile framework enables the rapid genetic modification of previously intractable microorganisms, expanding the synthetic biology toolbox for sustainable biomanufacturing and industrial biotechnology. ### Competing Interest Statement The authors have declared no competing interest. ## Funder Information Declared the Future Biomanufacturing Research Hub (FBRH), funded by the Engineering and Physical Sciences Research Council and the Biotechnology and Biological Sciences Research Council, (grant number EP/S01778X/1) the L'Oréal-UNESCO For Women in Science UK & Ireland Rising Talent grant Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a [CC-BY 4.0 International license](http://creativecommons.org/licenses/by/4.0/). bioRxiv and medRxiv thank the following for their generous financial support: > The Chan Zuckerberg Initiative, Cold Spring Harbor Laboratory, the Sergey Brin Family Foundation, California Institute of Technology, Centre National de la Recherche Scientifique, Fred Hutchinson Cancer Center, Imperial College London, Massachusetts Institute of Technology, Stanford University, The University of Edinburgh, University of Washington, and Vrije Universiteit Amsterdam. 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