Generalized Strategy for Engineering Mammalian Cell-Compatible RNA-Based Biosensors from Random Sequence Libraries.
ACS Sens
; 8(5): 2079-2086, 2023 05 26.
Article
en En
| MEDLINE
| ID: mdl-37155734
ABSTRACT
Fluorescent RNA-based biosensors are useful tools for real-time detection of molecules in living cells. These biosensors typically consist of a chromophore-binding aptamer and a target-binding aptamer, whereby the chromophore-binding aptamer is destabilized until a target is captured, which causes a conformational change to permit chromophore binding and an increase in fluorescence. The target-binding region is typically fabricated using known riboswitch motifs, which are already known to have target specificity and undergo structural changes upon binding. However, known riboswitches only exist for a limited number of molecules, significantly constraining biosensor design. To overcome this challenge, we designed a framework for producing mammalian cell-compatible biosensors using aptamers selected from a large random library by Capture-SELEX. As a proof-of-concept, we generated and characterized a fluorescent RNA biosensor against L-dopa, the precursor of several neurotransmitters. Overall, we suggest that this approach will have utility for generating RNA biosensors that can reliably detect custom targets in mammalian cells.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Técnicas Biosensibles
/
Aptámeros de Nucleótidos
Tipo de estudio:
Clinical_trials
Límite:
Animals
Idioma:
En
Revista:
ACS Sens
Año:
2023
Tipo del documento:
Article
País de afiliación:
Estados Unidos