High-contrast imaging of cellular non-repetitive drug-resistant genes via in situ dead Cas12a-labeled PCR.
Ruijie DengXinlei ZhangJijuan CaoXinmiao LiuYong ZhangFeng WangXuhan XiaPublished in: Chemical communications (Cambridge, England) (2024)
In situ imaging of genes of pathogenic bacteria can profile cellular heterogeneity, such as the emergence of drug resistance. Fluorescence in situ hybridization (FISH) serves as a classic approach to image mRNAs inside cells, but it remains challenging to elucidate genomic DNAs and relies on multiple fluorescently labeled probes. Herein, we present a dead Cas12a (dCas12a)-labeled polymerase chain reaction (CasPCR) assay for high-contrast imaging of cellular drug-resistant genes. We employed a syncretic dCas12a-green fluorescent protein (dCas12a-GFP) to tag the amplicons, thereby enabling high-contrast imaging and avoiding multiple fluorescently labeled probes. The CasPCR assay can quantify quinolone-resistant Salmonella enterica in mixed populations and identify them isolated from poultry farms.
Keyphrases
- drug resistant
- multidrug resistant
- high resolution
- acinetobacter baumannii
- pet imaging
- magnetic resonance
- fluorescence imaging
- crispr cas
- genome wide
- single molecule
- living cells
- small molecule
- genome editing
- induced apoptosis
- magnetic resonance imaging
- oxidative stress
- computed tomography
- deep learning
- high frequency
- dna methylation
- signaling pathway
- single cell
- cell death
- machine learning
- copy number
- transcription factor
- quantum dots
- photodynamic therapy
- energy transfer