RNA-triggered Cas12a2 antibacterial activity and resistance plasmid degradation
- 1China Pharmaceutical University
Abstract
Antimicrobial resistance (AMR) necessitates programmable antibacterial strategies with novel mechanisms. Here, we evaluated an inducible intracellular CRISPR/Cas12a2 system in engineered Escherichia coli targeting a chloramphenicol-resistance (Chl) transcript. Cas12a2 activation significantly delayed bacterial growth and reduced recoverable colony-forming units (CFU) by 3-log10 in vitro, and reduced bacterial burdens in a mouse skin wound model. Transcriptomic profiling revealed upregulation of recA, a key mediator of the SOS response. We therefore designed a Chl-recA dual-targeting construct, which produced a greater growth delay than either single-targeting counterpart. The optimized dual-targeting system led to an approximately 4-log10 reduction in bacterial burden in vivo. Furthermore, exploratory qPCR of culture supernatants suggested a treatment-associated decrease in amplifiable plasmid DNA. Overall, this study provides proof-of-concept for resistance-transcript-guided Cas12a2 antibacterial activity and highlights the potential of co-targeting stress-associated transcripts to enhance programmable antimicrobials.
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BibTeX
@misc{bai2026rnatriggered,
title = {{RNA-triggered Cas12a2 antibacterial activity and resistance plasmid degradation}},
author = {Y. Bai and Y. Song and W. Gu and H. Wu and Y. Liu},
howpublished = {bioRxiv},
year = {2026},
month = nov,
doi = {10.64898/2026.11.03.763947},
url = {https://www.biorxiv.org/content/10.64898/2026.11.03.763947v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - RNA-triggered Cas12a2 antibacterial activity and resistance plasmid degradation
AU - Y. Bai
AU - Y. Song
AU - W. Gu
AU - H. Wu
AU - Y. Liu
PY - 2026
DA - 2026/11/03
PB - bioRxiv
DO - 10.64898/2026.11.03.763947
UR - https://www.biorxiv.org/content/10.64898/2026.11.03.763947v1
AB - Antimicrobial resistance (AMR) necessitates programmable antibacterial strategies with novel mechanisms. Here, we evaluated an inducible intracellular CRISPR/Cas12a2 system in engineered Escherichia coli targeting a chloramphenicol-resistance (Chl) transcript. Cas12a2 activation significantly delayed bacterial growth and reduced recoverable colony-forming units (CFU) by 3-log10 in vitro, and reduced bacterial burdens in a mouse skin wound model. Transcriptomic profiling revealed upregulation of recA, a key mediator of the SOS response. We therefore designed a Chl-recA dual-targeting construct, which produced a greater growth delay than either single-targeting counterpart. The optimized dual-targeting system led to an approximately 4-log10 reduction in bacterial burden in vivo. Furthermore, exploratory qPCR of culture supernatants suggested a treatment-associated decrease in amplifiable plasmid DNA. Overall, this study provides proof-of-concept for resistance-transcript-guided Cas12a2 antibacterial activity and highlights the potential of co-targeting stress-associated transcripts to enhance programmable antimicrobials.
ER -