The CRISPR-dCas9 interference system suppresses inhA gene expression in Mycobacterium smegmatis.
Mycobacterium smegmatis
/ genetics
Bacterial Proteins
/ genetics
CRISPR-Cas Systems
Gene Expression Regulation, Bacterial
/ drug effects
Oxidoreductases
/ genetics
Isoniazid
/ pharmacology
Antitubercular Agents
/ pharmacology
Rifampin
/ pharmacology
Microbial Sensitivity Tests
Clustered Regularly Interspaced Short Palindromic Repeats
/ genetics
inhA
CRISPR interference
DCas9
Mycobacteria
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
30 10 2024
30 10 2024
Historique:
received:
28
06
2024
accepted:
22
10
2024
medline:
31
10
2024
pubmed:
31
10
2024
entrez:
31
10
2024
Statut:
epublish
Résumé
CRISPR-dead Cas9 interference (CRISPRi) has become a valuable tool for precise gene regulation. In this study, CRISPRi was designed to target the inhA gene of Mycobacterium smegmatis (Msm), a gene necessary for mycolic acid synthesis. Our findings revealed that sgRNA2 induced with 100 ng/ml aTc achieved over 90% downregulation of inhA gene expression and inhibited bacterial viability by approximately 1,000-fold. Furthermore, CRISPRi enhanced the susceptibility of M. smegmatis to isoniazid and rifampicin, which are both 50% and 90% lower than those of the wild-type strain or other strains, respectively. This study highlights the ability of CRISPRi to silence the inhA gene, which impacts bacterial viability and drug susceptibility. The findings provide valuable insights into the utility of CRISPRi as an alternative tool for gene regulation. CRISPRi might be further assessed for its synergistic effect with current anti-tuberculosis drugs and its possible implications for combating mycobacterial infections, especially drug-resistant tuberculosis.
Identifiants
pubmed: 39478003
doi: 10.1038/s41598-024-77442-2
pii: 10.1038/s41598-024-77442-2
doi:
Substances chimiques
Bacterial Proteins
0
InhA protein, Mycobacterium
EC 1.3.1.9
Oxidoreductases
EC 1.-
Isoniazid
V83O1VOZ8L
Antitubercular Agents
0
Rifampin
VJT6J7R4TR
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
26116Subventions
Organisme : Thailand Science Research and Innovation Fund of Chulalongkorn University
ID : CU_FRB65_hea (81)_176_37_06
Informations de copyright
© 2024. The Author(s).
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