Engineering Wax Ester Composition in Euglena gracilis Using Genome Editing.

Sakura Nagamine, Rikuto Oishi, Masami Nakazawa

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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4 · The record

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5 · Who and what money

Authors and funding

3 authors.

Sakura NagamineDepartment of Applied Biochemistry, Faculty of Agriculture, Osaka Metropolitan University, Sakai, Osaka, Japan.

Rikuto OishiDepartment of Applied Biochemistry, Faculty of Agriculture, Osaka Metropolitan University, Sakai, Osaka, Japan.

Masami NakazawaDepartment of Applied Biochemistry, Faculty of Agriculture, Osaka Metropolitan University, Sakai, Osaka, Japan. mami@omu.ac.jp.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Genome editing technologies have significantly expanded the potential for metabolic engineering in non-model organisms. In Euglena gracilis, genome editing methods using Cas9 and Cas12a were reported in 2019 and 2024, respectively, and are increasingly being applied to modify metabolic functions. This chapter provides a detailed protocol for CRISPR/Cas9-based genome editing that enables stable modification of wax ester composition under anaerobic conditions. By targeting key enzymes in the reversed β-oxidation pathway, the method allows the generation of knockout mutants with altered wax ester chain lengths. Beyond this application, the protocol supports reproducible and stable genetic modification of E. gracilis metabolism. It can be extended to the engineering of other biosynthetic pathways and is compatible with future integration of knock-in strategies. The approach offers a practical basis for the broader use of E. gracilis as a green chassis organism in synthetic biology and biomanufacturing.

Indexed as

EstersEuglena gracilisGene EditingMetabolic EngineeringWaxesCRISPR-Cas SystemsGene Knockout TechniquesEstersWaxesAnaerobic metabolismCRISPR/Cas9Euglena gracilisFatty acid β-oxidationGenome editingLoss-of-functionMetabolic modificationTargeted gene knockoutWax ester synthesis

Identifiers

PMID41627751

DOI10.1016/j.xpro.2021.101043

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.