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DOI RECORD

P. tricornutum CRISPR-Cas9 multiplexed mutant generation and purification v1

Florent Charton, Neo Revillon, Priscillia Pierre-Elies, Helena Cruz de Carvalho

DOI10.17504/protocols.io.kxygx3yyog8j/v1

PublisherSpringer Science and Business Media LLC

Journal / Source—

Published2023-08-30

Metadata Deposited2026-10-09 (updated: 2026-10-09)

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Typeposted-content

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Citations0

References deposited0

Access / license metadataOpen license identified License 1 ↗A reuse license does not by itself establish whether the full text is freely readable.

Abstract

Phaeodactylum tricornutum is a model diatom with a growing molecular toolbox available for functional genomics studies. This protocol provides all the information needed to generate biallelic mutants in P. tricornutum, harboring genomic loci ablations, using a single multiplexed CRISPR-Cas9 plasmid and bacterial conjugation. It can be used to knockout lncRNAs or protein coding genes. It can also be adapted to other diatoms by replacing the P. tricornutum promoter(s) by species specific ones.

Open DOI