Fluorescence‐based <scp>CRISPR</scp> interference system for controlled genetic repression and live single‐cell imaging in mycobacteria
In this research letter, we report the development and validation of a new subset of fluorescence-based CRISPR interference (CRISPRi) tools for our scientific community. The pJL series is directly derived from the original pIRL CRISPRi vectors and conserves all the elements to perform inducible targeted gene repression. These vectors carry two distinct fluorescent markers under the constitutive promoter psmyc to simplify the selection of recombinant clones. We demonstrate the functionality of these vectors by targeting the expression of the glycopeptidolipid translocase mmpL4b and the essential genes rpoB and mmpL3. Finally, we describe an efficient single-step procedure to co-transform mycobacterial species with this integrative genetic tool alongside episomal vectors. Such tools and approaches should be useful to foster discovery in mycobacterial research.
Authors
- Céline Crauste (ORCID: https://orcid.org/0000-0002-5714-8749)
- Stéphane Canaan (ORCID: https://orcid.org/0000-0001-7478-300X)
- Pierre Santucci (ORCID: https://orcid.org/0000-0002-6291-3425)
- Vanessa Point
- Wafaa Achache
- Janïs Laudouze
Institutions
- École Nationale Supérieure de Chimie de Montpellier (FR)
- Centre National de la Recherche Scientifique (FR)
- Université de Montpellier (FR)
- Aix-Marseille Université (FR)
- Méditerranée Infection Foundation (FR)
- Institut des Biomolécules Max Mousseron (FR)
Publication Details
- Journal
- FEBS Letters
- Published
- 2024-12-01
- DOI
- https://doi.org/10.1002/1873-3468.15071
- Citations
- 2
- Primary Topic
- Mycobacterium research and diagnosis
- Type
- article
- Field-Weighted Citation Impact
- 0.77
Funders
- Styrelsen för Internationellt Utvecklingssamarbete
- Aix-Marseille Université
- Agence Nationale de Recherches sur le Sida et les Hépatites Virales
- Centre National de la Recherche Scientifique
- Institut des sciences biologiques