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Destabilized reporters for background-subtracted, chemically-gated, and multiplexed deep-tissue imaging.


ABSTRACT: Tracking gene expression in deep tissues requires genetic reporters that can be unambiguously detected using tissue penetrant techniques. Magnetic resonance imaging (MRI) is uniquely suited for this purpose; however, there is a dearth of reporters that can be reliably linked to gene expression with minimal interference from background tissue signals. Here, we present a conceptually new method for generating background-subtracted, drug-gated, multiplex images of gene expression using MRI. Specifically, we engineered chemically erasable reporters consisting of a water channel, aquaporin-1, fused to destabilizing domains, which are stabilized by binding to cell-permeable small-molecule ligands. We showed that this approach allows for highly specific detection of gene expression through differential imaging. In addition, by engineering destabilized aquaporin-1 variants with orthogonal ligand requirements, it is possible to distinguish distinct subpopulations of cells in mixed cultures. Finally, we demonstrated this approach in a mouse tumor model through differential imaging of gene expression with minimal background.

SUBMITTER: Yun J 

PROVIDER: S-EPMC11253201 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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Destabilized reporters for background-subtracted, chemically-gated, and multiplexed deep-tissue imaging.

Yun Jason J   Huang Yimeng Y   Miller Austin D C ADC   Chang Brandon L BL   Baldini Logan L   Dhanabalan Kaamini M KM   Li Eugene E   Li Honghao H   Mukherjee Arnab A  

Chemical science 20240604 28


Tracking gene expression in deep tissues requires genetic reporters that can be unambiguously detected using tissue penetrant techniques. Magnetic resonance imaging (MRI) is uniquely suited for this purpose; however, there is a dearth of reporters that can be reliably linked to gene expression with minimal interference from background tissue signals. Here, we present a conceptually new method for generating background-subtracted, drug-gated, multiplex images of gene expression using MRI. Specifi  ...[more]

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