Work

Protocol to investigate gene expression heterogeneity in cyanobacteria using mRNA CARD-FISH

Year: 2025

Type: article

Abstract: Here, we present a protocol for visualizing gene expression in the filamentous cyanobacterium Trichodesmium and the single-celled species Synechocystis and Cyanothece using the catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) technique. We describe steps for fixation, agarose coating, enzymatic permeabilization, and sample handling. This protocol is broadly applicable to cyanobacteria, and the detection of rbcL mRNA in Trichodesmium, used as an example, supports its use to study the heterogeneity of physiological processes at single-cell level.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104212

Authors: Anxhela Hania, Louison Dufour, Ondřej Prášil, Kasia Piwosz, Meri Eichner

Institutions: University of South Bohemia in České Budějovice, Czech Academy of Sciences, Institute of Microbiology, National Marine Fisheries Research Institute

Language: English

Topic: Molecular Biology Techniques and Applications

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: Ministerstvo Školství, Mládeže a Tělovýchovy, Grantová Agentura České Republiky