| Autor: |
Weilong Shang, Zhen Hu, Mengyang Li, Yuting Wang, Yifan Rao, Li Tan, Juan Chen, Xiaonan Huang, Lu Liu, He Liu, Zuwen Guo, Huagang Peng, Yi Yang, Qiwen Hu, Shu Li, Xiaomei Hu, Jiao Zou, Xiancai Rao |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
mLife, Vol 2, Iss 4, Pp 462-478 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
2770-100X |
| DOI: |
10.1002/mlf2.12091 |
| Popis: |
Abstract Focal and systemic infections are serious threats to human health. Preclinical models enable the development of new drugs and therapeutic regimens. In vivo, animal bioluminescence (BL) imaging has been used with bacterial reporter strains to evaluate antimicrobial treatment effects. However, high‐sensitivity bioluminescent systems are required because of the limited tissue penetration and low brightness of the BL signals of existing approaches. Here, we report that NanoLuc (Nluc) showed better performance than LuxCDABE in bacteria. However, the retention rate of plasmid constructs in bacteria was low. To construct stable Staphylococcus aureus reporter strains, a partner protein enolase (Eno) was identified by screening of S. aureus strain USA300 for fusion expression of Nluc‐based luciferases, including Nluc, Teluc, and Antares2. Different substrates, such as hydrofurimazine (HFZ), furimazine (FUR), and diphenylterazine (DTZ), were used to optimize a stable reporter strain/substrate pair for BL imaging. S. aureus USA300/Eno‐Antares2/HFZ produced the highest number of photons of orange‐red light in vitro and enabled sensitive BL tracking of S. aureus in vivo, with sensitivities of approximately 10 CFU from mouse skin and 750 CFU from mouse kidneys. USA300/Eno‐Antares2/HFZ was a powerful combination based on the longitudinal evaluation of the therapeutic efficacy of antibiotics. The optimized S. aureus Eno‐Antares2/HFZ pair provides a technological advancement for the in vivo evaluation of antimicrobial treatment. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
|
Plný text