CRISPR Interference-Based Inhibition of MAB_0055c Expression Alters Drug Sensitivity in Mycobacterium abscessus

Abstract

In this study, we utilized CRISPR interference (CRISPRi) to specifically target the MAB_0055c gene in M. abscessus , a bacterium that causes difficult-to-treat infections. The study found that silencing the gene lead to increased rifabutin and rifalazil susceptibility.

Ceclazepide as novelMycobacterium abscessusinhibitor

“…Additionally, factors such as aging, underlying pulmonary conditions like cystic fibrosis (CF), and immunocompromised states such as acquired immunodeficiency syndrome (AIDS) contribute to the heightened risk of MAC and M. abscessus infections in numerous regions worldwide (Johansen et al, 2020). M. abscessus infections pose a significant challenge, especially due to their high resistance levels (Nguyen et al, 2024). The clinical manifestations of M. abscessus infection are typically classified as either pulmonary or extrapulmonary disease.…”

“…Despite being less harmful to humans when compared to M. tuberculosis , the bacterium responsible for causing tuberculosis, treating lung infections caused by M. abscessus is extremely challenging. This is primarily due to the bacterium’s high resistance to commonly used antibiotics, such as β-lactams, tetracyclines, aminoglycosides, and macrolides (Luthra et al, 2018; Nguyen et al, 2024). Additionally, even anti-tuberculosis drugs like rifampicin and isoniazid prove to be ineffective against M. abscessus .…”

“…A prime example of this is the comparison between rifampicin and rifabutin, another rifamycin analog. Rifabutin has demonstrated remarkable activity against M. abscessus both in vivo and in vitro (Aziz et al, 2017; Nguyen et al, 2023a). This can be attributed to its favorable oral bioavailability, excellent pharmacokinetic properties (including a long half-life and high cellular penetration), and its ability to achieve appropriate concentrations in human lung tissue.…”

“…In recent times, a number of newly developed chemical entities and antimicrobial compounds have emerged, displaying promising activity against the highly drug-resistant pathogen M. abscessus . Some notable examples include ohmyungsamycin, omadacycline, epetraborole, DS86760016, delpazolid, etc (Cho et al, n.d.; Kim et al, n.d.; Aziz et al, 2017; Bich Hanh et al, 2021; Egorova et al, 2021; Ganapathy et al, 2021; Jeon et al, 2022; Fujiwara et al, 2023; Nguyen et al, 2023b, 2023a). These compounds offer potential avenues for combating the challenges posed by M. abscessus infections.…”

Investigating Mtr and MshC as potential targets in the fight against Mycobacterium abscessus infections

“…Springer et al observed that excision of the integrated plasmid is by the integrase and occurs more frequently than anticipated, but can be avoided by utilizing a second nonreplicating Int-containing plasmid Furthermore, integrase-mediated excision of the L5 integrated plasmid can lead to plasmid loss in cases where the integrated plasmid is disadvantageous for the growth of the recombinant strain (195), indicating that the pLJR965 plasmid may be toxic for Mab over a longer period of time. Nevertheless, any loss of the integrated plasmid was not reported when applying the Sth CRISPRi-system on either Msm, Mtb or Mab (146,148,196,197), yet Bosch et al described the use of a suicide plasmid to express Int in order to enhance the stability of the L5 integration during long-term studies (191). It is unclear whether excision of the integrated plasmid occurs more frequently in Mab than in Msm or Mtb, we waited too long in between experiments leading to the loss of the unstable plasmid over time, or if we were unable to correctly translate the Sth CRISPRi-system for use in…”