Efficient CRISPR-Cas genome editing in brown algae

Cláudia Martinho; Masakazu Hoshino; Morgane Raphalen; Viktoriia Bukhanets; Anagha Kerur; Kenny A. Bogaert; Rémy Luthringer; Susana M. Coelho

doi:10.1016/j.crmeth.2025.101273

Efficient CRISPR-Cas genome editing in brown algae

Cláudia Martinho (Lead / Corresponding author)
, Masakazu Hoshino (Lead / Corresponding author)
, Morgane Raphalen
, Viktoriia Bukhanets
, Anagha Kerur
, Kenny A. Bogaert
, Rémy Luthringer
, Susana M. Coelho (Lead / Corresponding author)

Plant Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

Brown algae represent the third most complex lineage to have independently evolved multicellularity, distinct from plants and animals. Yet, functional studies of their development and evolution have been limited by the absence of efficient genome editing tools. Here, we present a robust, high-efficiency, and transgene-free CRISPR-based genome editing platform applicable across four ecologically and biotechnologically important brown algal species. Using Ectocarpus as a model, we optimized a polyethylene glycol (PEG)-mediated ribonucleoprotein (RNP) delivery system that achieves reproducible editing across multiple loci without cloning or specialized equipment. As proof of concept, we recreated the hallmark imm mutant phenotype by precisely editing the IMMEDIATE UPRIGHT (IMM) locus. APT/2-fluoroadenine (2-FA) selection further enhanced specificity with minimal false positives. The method was easily transferable to other species, including kelps. This platform now enables functional genomics in brown algae, providing powerful tools for investigating development, life cycle regulation, and the independent evolution of complex multicellularity.

Original language	English
Article number	101273
Journal	Cell Reports Methods
Volume	6
Issue number	1
DOIs	https://doi.org/10.1016/j.crmeth.2025.101273
Publication status	Published - 30 Dec 2025

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Final published version
Martinho et al., 2026, Cell Reports Methods 6, 101273 January 26, 2026 © 2025 The Author(s). Published by Elsevier Inc. https://doi.org/10.1016/j.crmeth.2025.101273
Final published version, 5.52 MBLicence: CC BY

Cite this

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title = "Efficient CRISPR-Cas genome editing in brown algae",

abstract = "Brown algae represent the third most complex lineage to have independently evolved multicellularity, distinct from plants and animals. Yet, functional studies of their development and evolution have been limited by the absence of efficient genome editing tools. Here, we present a robust, high-efficiency, and transgene-free CRISPR-based genome editing platform applicable across four ecologically and biotechnologically important brown algal species. Using Ectocarpus as a model, we optimized a polyethylene glycol (PEG)-mediated ribonucleoprotein (RNP) delivery system that achieves reproducible editing across multiple loci without cloning or specialized equipment. As proof of concept, we recreated the hallmark imm mutant phenotype by precisely editing the IMMEDIATE UPRIGHT (IMM) locus. APT/2-fluoroadenine (2-FA) selection further enhanced specificity with minimal false positives. The method was easily transferable to other species, including kelps. This platform now enables functional genomics in brown algae, providing powerful tools for investigating development, life cycle regulation, and the independent evolution of complex multicellularity.",

author = "Cl{\'a}udia Martinho and Masakazu Hoshino and Morgane Raphalen and Viktoriia Bukhanets and Anagha Kerur and Bogaert, \{Kenny A.\} and R{\'e}my Luthringer and Coelho, \{Susana M.\}",

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AU - Martinho, Cláudia

AU - Hoshino, Masakazu

AU - Raphalen, Morgane

AU - Bukhanets, Viktoriia

AU - Kerur, Anagha

AU - Bogaert, Kenny A.

AU - Luthringer, Rémy

AU - Coelho, Susana M.

PY - 2025/12/30

Y1 - 2025/12/30

N2 - Brown algae represent the third most complex lineage to have independently evolved multicellularity, distinct from plants and animals. Yet, functional studies of their development and evolution have been limited by the absence of efficient genome editing tools. Here, we present a robust, high-efficiency, and transgene-free CRISPR-based genome editing platform applicable across four ecologically and biotechnologically important brown algal species. Using Ectocarpus as a model, we optimized a polyethylene glycol (PEG)-mediated ribonucleoprotein (RNP) delivery system that achieves reproducible editing across multiple loci without cloning or specialized equipment. As proof of concept, we recreated the hallmark imm mutant phenotype by precisely editing the IMMEDIATE UPRIGHT (IMM) locus. APT/2-fluoroadenine (2-FA) selection further enhanced specificity with minimal false positives. The method was easily transferable to other species, including kelps. This platform now enables functional genomics in brown algae, providing powerful tools for investigating development, life cycle regulation, and the independent evolution of complex multicellularity.

AB - Brown algae represent the third most complex lineage to have independently evolved multicellularity, distinct from plants and animals. Yet, functional studies of their development and evolution have been limited by the absence of efficient genome editing tools. Here, we present a robust, high-efficiency, and transgene-free CRISPR-based genome editing platform applicable across four ecologically and biotechnologically important brown algal species. Using Ectocarpus as a model, we optimized a polyethylene glycol (PEG)-mediated ribonucleoprotein (RNP) delivery system that achieves reproducible editing across multiple loci without cloning or specialized equipment. As proof of concept, we recreated the hallmark imm mutant phenotype by precisely editing the IMMEDIATE UPRIGHT (IMM) locus. APT/2-fluoroadenine (2-FA) selection further enhanced specificity with minimal false positives. The method was easily transferable to other species, including kelps. This platform now enables functional genomics in brown algae, providing powerful tools for investigating development, life cycle regulation, and the independent evolution of complex multicellularity.

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Efficient CRISPR-Cas genome editing in brown algae

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